Publication Information

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3930, 3957, 3983, 3988, 4011, 4031, 4054, 4064, 4091, 4124, 4128, 4155, 4204, 4219, 4256, 4313], 'best': [21, 285, 4067], 'method': [22, 286], 'evolve': [24, 288], 'protein': [25, 289, 1082, 1288, 3200, 3232, 3239, 3250], 'molecular': [26, 290, 546], 'recognition.': [27, 291], 'We': [28, 292, 1091, 1375, 1388, 1466, 1591, 1834, 2757, 3499, 3541, 3679, 3709, 3922], 'previously': [29, 293, 1377, 1891], 'used': [30, 294, 878, 906, 1166, 2595, 3710], 'DNA': [31, 151, 229, 295, 415, 493, 1421, 2192, 2228, 2246, 2252, 2267, 2275, 2313, 2324, 2601, 2610, 2645, 2738, 2784, 3126], 'shuffling': [32, 230, 296, 494, 1422, 1871, 2739], 'direct': [34, 298, 1581, 3454], 'of': [37, 60, 89, 93, 109, 122, 134, 163, 175, 178, 199, 203, 214, 226, 245, 257, 301, 324, 353, 357, 373, 386, 398, 427, 439, 442, 463, 467, 478, 490, 509, 521, 545, 552, 565, 581, 673, 704, 709, 867, 948, 1075, 1081, 1087, 1097, 1106, 1115, 1117, 1124, 1137, 1170, 1279, 1296, 1363, 1381, 1392, 1420, 1470, 1584, 1774, 1797, 1811, 1838, 1844, 1860, 1864, 1881, 1980, 2033, 2044, 2080, 2102, 2189, 2226, 2238, 2245, 2250, 2256, 2265, 2273, 2474, 2483, 2521, 2528, 2542, 2599, 2606, 2624, 2654, 2675, 2696, 2765, 2849, 2951, 2960, 3016, 3152, 3247, 3257, 3277, 3323, 3339, 3345, 3457, 3510, 3563, 3728, 3735, 3741, 3810, 3841, 3856, 3897, 3908, 3929, 3968, 3982, 3987, 4004, 4013, 4022, 4053, 4063, 4090, 4130, 4136, 4142, 4146, 4158, 4188, 4206, 4247, 4259, 4289, 4304, 4316], 'Escherichia': [38, 302, 1268], 'coli': [39, 303, 1269, 1299, 1337, 1400, 2342, 2628, 2919, 3106, 3650, 3654], 'β-glucuronidase': [40, 304, 874, 885, 890, 913, 918, 1338], '(GUS)': [41, 305], 'variants': [42, 306, 1119, 1383, 1586, 3459], 'with': [43, 116, 149, 193, 195, 307, 380, 413, 457, 459, 554, 706, 1307, 1369, 1384, 1587, 2061, 2144, 2182, 2195, 2262, 2414, 2448, 2564, 2633, 2703, 2762, 2804, 2882, 2924, 2976, 3023, 3035, 3048, 3075, 3111, 3122, 3254, 3460, 3753, 3756, 3768, 3854, 3875, 3979, 3992, 4096, 4201], 'increased': [44, 112, 308, 376, 1385, 1404, 1588, 2936], 'β-galactosidase': [45, 309, 1386, 1405, 1451, 4101], 'activity.': [46, 310, 1387, 1590, 2938, 3125, 3462, 3904], 'Epistatic': [47, 311], '(synergistic)': [48, 312], 'mutations': [49, 313, 1139, 1472, 3937, 4035], 'in': [50, 67, 97, 114, 166, 171, 190, 237, 314, 331, 361, 378, 430, 435, 454, 501, 587, 677, 954, 968, 1104, 1120, 1140, 1176, 1357, 1367, 1397, 1450, 1483, 1496, 1598, 1770, 1804, 1818, 1828, 1876, 2070, 2186, 2277, 2383, 2435, 2452, 2468, 2480, 2487, 2506, 2538, 2576, 2650, 2660, 2768, 2788, 2798, 2814, 2855, 2898, 2947, 2968, 3062, 3087, 3176, 3267, 3283, 3329, 3415, 3427, 3520, 3547, 3684, 3738, 3775, 3814, 3862, 3938, 3949, 3959, 3973, 3977, 4072, 4100, 4123, 4139, 4154, 4166, 4212, 4255, 4267, 4312, 4324], 'amino': [51, 315, 726, 971, 1293, 1480, 1951, 3545], 'acids': [52, 316, 1481, 1952, 3546], '557,': [53, 317, 1953, 3513], '566,': [54, 318, 1954, 3514], 'and': [55, 72, 100, 153, 231, 248, 319, 336, 364, 417, 495, 512, 575, 583, 684, 773, 799, 829, 935, 1059, 1099, 1129, 1300, 1371, 1444, 1459, 1476, 1786, 1790, 1854, 1936, 1955, 1982, 2008, 2016, 2052, 2065, 2086, 2128, 2138, 2147, 2204, 2247, 2269, 2299, 2323, 2381, 2433, 2464, 2491, 2510, 2547, 2555, 2568, 2581, 2614, 2618, 2637, 2658, 2671, 2693, 2706, 2716, 2734, 2791, 2822, 2847, 2886, 2893, 2906, 2916, 2928, 2955, 3027, 3039, 3043, 3051, 3060, 3069, 3078, 3082, 3095, 3103, 3115, 3145, 3172, 3192, 3197, 3208, 3214, 3274, 3429, 3515, 3622, 3646, 3656, 3772, 3793, 3807, 4078, 4182, 4216, 4283], '568,': [56, 320, 3516], 'which': [57, 321, 3517, 3556], 'are': [58, 322, 830, 974, 985, 1304], 'part': [59, 323, 2848], 'an': [61, 161, 325, 425, 1484, 3063, 3204, 3337, 3733, 3776], 'active': [62, 326, 982, 1310, 1485, 3940], 'site': [63, 86, 327, 350, 983, 1486, 3503, 3941], 'loop,': [64, 328], 'were': [65, 127, 142, 169, 212, 329, 391, 406, 433, 476, 1414, 1453, 1457, 1596, 1780, 1890, 1916, 1957, 2059, 2089, 2260, 2345, 2375, 2427, 2455, 2462, 2517, 2544, 2640, 2656, 2796, 2812, 2853, 2896, 2931, 2953, 2965, 2982, 3084, 3109, 3118, 3131, 3174, 3216, 3251, 3290, 3518, 3751, 3765, 3797, 3805, 3820, 3827, 3923, 3932, 3947, 3962, 3971, 4047], 'identified': [66, 330, 1597, 3519], 'that': [68, 85, 133, 210, 218, 250, 332, 349, 397, 474, 482, 514, 560, 711, 951, 1478, 1595, 1847, 2527, 2589, 2593, 2973, 2988, 3726, 3834, 3869, 3883, 3956, 4036, 4062, 4087, 4175, 4276, 4333], 'experiment': [69, 333, 3525], '(Matsumura,': [70, 334], 'I.,': [71, 335], 'Ellington,': [73, 337], 'A.': [74, 338, 607, 820, 857, 1435, 1720, 1724, 1999, 2748, 2873, 3607, 3620], 'D.': [75, 339, 1550], '(2001)': [76, 340], 'J.': [77, 341, 613, 640, 1003, 1016, 1151, 1503, 1535, 1548, 1569, 1618, 1683, 1704, 1732, 1901, 3488, 3530, 4108], 'Mol.': [78, 342, 596, 1017, 1152, 1504, 1570, 1619, 1705, 1902, 3489, 3531], 'Biol.': [79, 343, 642, 737, 782, 1018, 1041, 1153, 1230, 1326, 1505, 1706, 1903, 3490, 3532], '305,': [80, 344], '331–339).': [81, 345], 'Here': [82, 346, 1514], 'we': [83, 347, 1515, 2241, 2594, 4085], 'show': [84, 348], 'saturation': [87, 351, 1606, 2467, 3504, 3789, 4132], 'mutagenesis': [88, 352, 1475, 1520, 1607, 1785, 3505, 4089, 4133], 'these': [90, 354, 1471, 3467, 3791], 'residues,': [91, 355], 'overexpression': [92, 356], 'resulting': [95, 359, 1102, 2723, 2909, 3098, 3113], 'library': [96, 360, 1391, 1801, 3099, 3549, 3567], 'E.': [98, 156, 362, 420, 609, 1298, 1336, 1358, 2341, 2627, 2918, 3105, 3649, 3653], 'coli,': [99, 363, 1359], 'high': [101, 365, 1130, 1355, 1791, 2248, 2385, 2449, 3687, 3801], 'throughput': [102, 366, 1109, 1131, 1792, 2386, 3562, 3688], 'screening': [103, 367, 1110, 1132, 1793, 4159, 4260, 4317], 'led': [104, 368], 'rapid': [107, 371], 'clones': [110, 126, 159, 374, 390, 423, 1142, 1446, 1456, 1779, 1846, 2934, 2945, 3121, 3931, 3946, 4025, 4046, 4057], 'exhibiting': [111, 160, 375, 424, 1403, 1447, 2935], 'activity': [113, 168, 377, 432, 1406, 1452, 1859, 2439, 2451, 2520, 3666, 3852, 3872, 3915, 3961, 4003, 4052, 4102], 'reactions': [115, 379, 1368, 2029, 2058, 2279], 'p-nitrophenyl-β-d-xylopyranoside': [117, 381], '(pNP-xyl).': [118, 382], 'The': [119, 138, 383, 402, 690, 746, 1267, 1335, 1777, 1800, 1813, 1912, 1928, 2026, 2057, 2117, 2133, 2163, 2191, 2224, 2311, 2372, 2519, 2558, 2583, 2699, 2722, 2783, 2794, 2808, 2843, 2908, 3014, 3055, 3071, 3080, 3097, 3150, 3230, 3286, 3463, 3566, 3640, 3762, 3785, 3803, 3825, 3850, 3905, 3943, 4002, 4016, 4049, 4066, 4120, 4302], 'xylosidase': [120, 167, 384, 431, 1832, 1858, 2450, 3960], 'activities': [121, 385, 1366], '14': [124, 139, 388, 403, 2535, 4056], 'fittest': [125, 389, 1461, 3945], '30-fold': [128, 392, 4059], 'higher': [129, 393, 4060, 4121], 'on': [130, 394, 725, 1407], 'average': [131, 162, 395, 426, 1814, 1843, 2512, 3338, 3734, 4050], 'than': [132, 396, 3879, 3916, 4061], 'wild-type': [136, 400], 'GUS.': [137, 401], 'corresponding': [140, 404, 4032], 'plasmids': [141, 405, 2536, 4033], 'pooled,': [143, 407, 2546], 'amplified': [144, 408, 2659, 2813, 2854, 2966, 3022, 3034, 3047, 3061, 3074], 'by': [145, 409, 1605, 1856, 1918, 1959, 2099, 2179, 2218, 2327, 2347, 2351, 2737, 2991, 3222, 3292, 3570, 4009, 4020, 4197, 4209, 4218], 'long': [146, 410, 2027, 2857, 2900, 3089], 'PCR,': [147, 411], 'self-ligated': [148, 412], 'T4': [150, 414, 2251, 2266, 2274, 2312, 2600, 2609], 'ligase,': [152, 416], 'transformed': [154, 418, 2346, 2373, 2632, 2923, 3110, 3647, 3716, 3755], 'into': [155, 419, 578, 721, 2378, 3648, 3718, 3838], 'coli.': [157, 421], 'Thirteen': [158, 422], '80-fold': [164, 428], 'improvement': [165, 185, 429, 449, 1849, 4017, 4071], 'isolated': [170, 434, 2649, 2946, 4138, 4153, 4254, 4311], 'second': [173, 437, 2652], 'round': [174, 206, 438, 470, 1773, 1810, 2541, 2653, 2950, 4141, 4157, 4258, 4315], 'screening.': [176, 440], 'One': [177, 441], 'evolved': [180, 444, 1061, 1782, 3128, 3154, 3347, 4177, 4278, 4335], 'proteins': [181, 445, 3796], 'exhibited': [182, 446, 1848, 3912, 4019], '∼200-fold': [184, 448], 'over': [186, 450, 1820, 1850], 'wild': [188, 452, 1862, 3352, 3409, 4222], 'type': [189, 453, 3353], 'reactivity': [191, 455], '(kcat/Km)': [192, 456], 'pNP-xyl,': [194, 458, 896, 924], '290,000-fold': [197, 461], 'inversion': [198, 462], 'specificity.': [200, 464, 969], 'Sequence': [201, 465], 'analysis': [202, 466], '13': [205, 469, 1464], '2': [207, 471, 2720, 2778], 'isolates': [208, 472], 'suggested': [209, 473], 'all': [211, 475, 3543, 3870, 3928], 'products': [213, 477, 2811, 3058], 'intermolecular': [215, 479], 'recombination': [216, 480, 1789], 'events': [217, 481], 'occurred': [219, 483], 'during': [220, 484], 'whole': [221, 260, 485, 524, 1830, 1960, 2437, 3571, 4092], 'plasmid': [222, 261, 486, 525, 1961, 2635, 2926, 3465, 3572], 'PCR.': [223, 487], 'Further': [224, 488], 'rounds': [225, 489, 1419], 'using': [228, 492, 1977, 2168, 2211, 2407, 2552, 2664, 2824, 2903, 3066, 3092, 3133, 3159, 3179, 3225, 3236, 3434, 3627], 'staggered': [232, 496, 899, 927], 'extension': [233, 497, 900, 928, 2980], 'process': [234, 498, 676], '(StEP)': [235, 499], 'resulted': [236, 500], 'modest': [238, 502], 'improvement.': [239, 503], 'These': [240, 504, 716, 942, 4044], 'results': [241, 505], 'underscore': [242, 506], 'importance': [244, 508], 'epistatic': [246, 510], 'interactions': [247, 511], 'demonstrate': [249, 513, 3955], 'they': [251, 515], 'can': [252, 516, 1351, 2989], 'be': [253, 517, 665, 1352], 'optimized': [254, 518], 'through': [255, 519, 572, 669, 1473, 1783], 'variations': [256, 520], 'facile': [259, 523], 'PCR': [262, 526, 1962, 2028, 2140, 2164, 2172, 2207, 2258, 2663, 2724, 2732, 2760, 2817, 2826, 2858, 2902, 2911, 2970, 3065, 3091, 3573, 3641], 'technique.': [263, 527], 'Adaptive': [528], 'is': [530, 1052, 1068, 1271, 1343, 2022, 3336, 3557, 3635, 4170, 4183, 4271, 4284, 4328], 'arguably': [531], 'most': [533, 3902], 'fundamental': [534], 'biological': [535], 'process,': [536], 'although': [537], 'it': [538, 1350], 'remains': [539], 'poorly': [540], 'understood.': [541], 'A': [542, 3890], 'better': [543], 'understanding': [544, 1071], 'adaptation': [547], 'would': [548, 680], 'facilitate': [549], 'engineering': [551], 'enzymes': [553, 710, 717, 793, 1056, 1171, 2566, 3210], 'novel': [555], 'specificities.': [556], 'Theoreticians': [557], 'proposed': [559], 'relatively': [562], 'small': [563], 'number': [564], 'catalytically': [566], 'inefficient,': [567], 'broad': [568], 'specificity': [569, 1174, 1498], 'proto-enzymes': [570], 'diversified': [571], 'gene': [573, 1339, 2852, 3019, 3502, 4093], 'duplication': [574], 'adaptive': [576, 1280], 'multitude': [580, 708], 'efficient': [582], 'specialized': [584], 'catalysts': [585], 'present': [586], 'contemporary': [589], 'biosphere': [590], '(2Kacser': [591], 'H.': [592, 1196, 1672, 2995], 'Beeby': [593], 'R.': [594, 1722, 1992, 2866], 'J': [595], 'Evol.': [597, 1571, 1620], '1984;': [598], '20:': [599, 1652, 1736], '38-51Crossref': [600], 'PubMed': [601, 620, 633, 646, 659, 741, 786, 824, 1022, 1045, 1157, 1190, 1207, 1218, 1234, 1262, 1330, 1439, 1509, 1540, 1560, 1575, 1624, 1654, 1690, 1710, 1738, 1760, 1907, 1972, 2003, 2367, 2402, 2752, 2838, 2877, 3009, 3317, 3494, 3536, 3583, 3601, 3614, 3674, 3704], 'Scopus': [602, 621, 634, 647, 742, 787, 825, 1023, 1046, 1158, 1191, 1208, 1219, 1235, 1263, 1331, 1440, 1510, 1541, 1561, 1576, 1625, 1655, 1691, 1711, 1739, 1761, 1908, 1973, 2004, 2368, 2403, 2753, 2839, 2878, 3010, 3318, 3495, 3537, 3584, 3602, 3615, 3675, 3705], '(113)': [603], 'Google': [604, 623, 636, 649, 660, 744, 789, 827, 861, 1025, 1048, 1160, 1193, 1210, 1221, 1237, 1265, 1333, 1442, 1512, 1543, 1563, 1578, 1627, 1657, 1693, 1713, 1741, 1763, 1910, 1975, 2006, 2337, 2370, 2405, 2755, 2841, 2880, 3012, 3320, 3497, 3539, 3586, 3604, 3617, 3677, 3707], 'Scholar,': [605, 624, 637, 650, 1026, 1194, 1211, 1222, 1238, 1544, 1564, 1628, 1658, 1694, 1714, 1742, 3587, 3605], '3Lazcano': [606], 'Diaz-Villagomez': [608], 'Mills': [610], 'T.': [611, 1181], 'Oro': [612], 'Adv.': [614], 'Space': [615], 'Res.': [616, 2363], '1995;': [617, 821], '15:': [618, 1558], '345-356Crossref': [619], '(19)': [622], '4Jensen': [625], 'R.A.': [626], 'Annu.': [627], 'Rev.': [628], 'Microbiol.': [629, 3670], '1976;': [630], '30:': [631, 657], '409-425Crossref': [632], '(817)': [635], '5Ycas': [638], 'M.': [639, 1005, 1554, 2396, 3698], 'Theor.': [641], '1974;': [643], '44:': [644], '145-160Crossref': [645], '(160)': [648], '6Waley': [651], 'S.G.': [652, 1013], 'Comp.': [653], 'Biochem.': [654, 1536], 'Physiol.': [655], '1969;': [656], '1-11Crossref': [658], 'Scholar).': [661, 745, 790, 862, 1049, 1161, 1266, 1334, 1513, 1764, 1911, 2338, 2371, 2756, 2842, 3013, 3321, 3498, 3540, 3678, 3708, 4119], 'This': [662, 1765, 2984], 'supposition': [663], 'cannot': [664], 'proven': [666], 'nor': [667], 'refuted': [668], 'experimentation,': [670], 'recapitulation': [672], 'diversification': [675], 'laboratory': [679], 'support': [681, 4127], 'its': [682, 1851, 3657, 4167, 4268, 4325], 'feasibility': [683], 'reveal': [685], 'possible': [686], 'underlying': [687], 'structural': [688, 702, 1073], 'mechanisms.': [689], 'glycoside': [691, 871], 'hydrolases': [692], 'exemplify': [693], 'enzyme': [694, 1076, 2197], 'diversification.': [695], 'Natural': [696], 'selection': [697, 1815, 1867, 4191], 'has': [698, 1164, 1289], 'matched': [699], 'great': [701], 'diversity': [703], 'carbohydrates': [705], 'selectively': [712], 'catalyze': [713, 945], 'their': [714, 955, 3177], 'cleavage.': [715], 'been': [719, 1165, 1290], 'classified': [720], '91': [722], 'families': [723], 'based': [724], 'acid': [727, 972, 1294], 'sequence': [728, 1118], 'similarity': [729], '(7Bourne': [730], 'Y.': [731], 'Henrissat': [732], 'B.': [733, 776, 803, 840, 997, 1546, 1664, 2830], 'Curr.': [734, 779, 1184, 1201, 1227, 3669], 'Opin.': [735, 780, 1185, 1202, 1228], 'Struct.': [736, 781, 1040, 1325], '2001;': [738, 1187, 1215, 1259, 1506, 3533], '11:': [739], '593-600Crossref': [740], '(359)': [743], 'GH-A': [747, 792], 'clan': [748], '(sometimes': [749], 'called': [750, 1609], '4/7': [752], 'superfamily)': [753], 'comprises': [754], '>3000': [755], 'GenBank™': [756], 'sequences': [757, 973, 1295, 3509], 'from': [758, 835, 1926, 1940, 2944, 3261], 'Families': [759], '1,': [760], '2,': [761], '5,': [762], '10,': [763], '17,': [764], '26,': [765], '35,': [766], '39,': [767], '42,': [768], '53,': [769], '59,': [770], '72,': [771], '79,': [772], '86': [774], '(8Henrissat': [775], 'Davies': [777, 812, 849], 'G.': [778, 813, 850, 1660, 1670, 2025], '1997;': [783, 1968, 3579, 3671], '7:': [784], '637-644Crossref': [785], '(1400)': [788], 'All': [791, 1455], 'retain': [794], 'same': [796], '(β/α)8-barrel': [797], 'fold': [798], 'catalytic': [800, 1365], 'mechanism': [801], '(9Henrissat': [802], 'Callebaut': [804, 841], 'I.': [805, 842, 1150, 1500, 1900, 2395, 3303, 3487, 3527, 3623, 3697, 4104], 'Fabrega': [806, 843], 'S.': [807, 819, 844, 856, 1028, 1313, 1434, 1718, 1728, 1986, 1998, 2747, 2860, 2872], 'Lehn': [808, 845], 'P.': [809, 846, 1678], 'Mornon': [810, 847], 'J.P.': [811, 848], 'Proc.': [814, 851, 1429, 1993, 2742, 2867], 'Natl.': [815, 852, 1430, 1994, 2743, 2868], 'Acad.': [816, 853, 1431, 1995, 2744, 2869], 'Sci.': [817, 854, 1432, 1996, 2745, 2870], 'U.': [818, 855, 1433, 1997, 2746, 2871], '92:': [822], '7090-7094Crossref': [823], '(512)': [826], 'Scholar)': [828, 1443, 1579, 1976, 2007, 2406, 2881, 3618], 'thought': [831], 'diverged': [834], 'common': [837], 'ancestor': [838], '(10Henrissat': [839], '1996;': [858, 1042, 1327, 1707, 2834], '93:': [859], '5674PubMed': [860], 'Consider': [863], 'specific': [865], 'example': [866], 'two': [868, 1417], 'well': [869, 3722], 'characterized': [870, 2434, 3948], 'hydrolases:': [872], 'human': [873, 1286, 1301], '(GUS,': [875], '1The': [876], 'abbreviations': [877, 905], 'are:': [879, 907], 'GUS,': [880, 908], 'β-glucuronidase;': [881, 909], 'XYL,': [882, 910], 'β-xylosidase;': [883, 911], 'PSQ,': [884, 912], 'mutants': [886, 891, 914, 919, 2892, 3426, 3683, 3911, 4095, 4137], 'containing': [887, 892, 915, 920, 1411, 2280, 3550, 3629, 3900], 'Pro557/Ser566/Gln568;': [888, 916], 'PAF,': [889, 917], 'Pro557/Ala566/Phe568;': [893, 921], 'pNP-gluc,': [894, 922], 'p-nitrophenyl-β-d-glucuronide;': [895, 923], 'p-nitrophenyl-β-d-xylopyranoside;': [897, 925], 'StEP,': [898, 926], 'process;': [901, 929], 'LB,': [902, 930], 'Luria': [903, 931], 'broth.1The': [904], 'broth.': [932], 'Family': [933, 940], '2)': [934, 1915], 'Thermoanaerobacterium': [936], 'saccharolyticum': [937], 'β-xylosidase': [938], '(XYL,': [939], '39).': [941], 'distant': [943], 'homologs': [944, 1303], 'hydrolysis': [947], 'similar': [949, 2587], 'substrates': [950], 'differ': [952], 'only': [953, 4097], 'C5': [956], 'substituents': [957], '(carboxylate': [958], 'for': [959, 962, 1276, 1493, 1808, 1823, 2074, 2095, 2125, 2155, 2305, 2320, 2499, 2713, 2719, 2777, 2933, 3120, 3143, 3466, 3476, 3779, 3847, 3996], 'β-glucuronides,': [960], 'hydrogen': [961], 'β-xylosides),': [963], 'overlap': [965, 2825, 2901, 3064, 3090], 'very': [966], 'little': [967], 'Their': [970], 'too': [975], 'divergent': [976], 'align,': [978], 'six': [980], 'conserved': [981, 1309], 'residues': [984, 1594], 'superimposable': [986], '(Fig.': [987, 1914, 3920], '1)': [988], '(11Yang': [989], 'J.K.': [990], 'Yoon': [991], 'H.J.': [992, 994], 'Ahn': [993], 'Il': [995], 'Lee': [996], 'Pedelacq': [998], 'J.D.': [999], 'Liong': [1000], 'E.C.': [1001, 1730], 'Berendzen': [1002], 'Laivenieks': [1004], 'Vieille': [1006], 'C.': [1007, 1213, 1988, 2862], 'Zeikus': [1008], 'G.J.': [1009], 'Vocadlo': [1010], 'D.J.': [1011, 1648], 'Withers': [1012], 'Suh': [1014], 'S.W.': [1015, 1242], '2004;': [1019, 2399, 3701], '335:': [1020], '155-165Crossref': [1021], '(63)': [1024, 1762], '12Jain': [1027], 'Drendel': [1029, 1314], 'W.B.': [1030, 1315], 'Chen': [1031, 1199, 1316, 1677], 'Z.W.': [1032, 1317], 'Mathews': [1033, 1318], 'F.S.': [1034, 1319], 'Sly': [1035, 1320], 'W.S.': [1036, 1321], 'Grubb': [1037, 1322], 'J.H.': [1038, 1323], 'Nat.': [1039, 1324, 1649, 1733, 3004, 3312], '3:': [1043, 1328], '375-381Crossref': [1044, 1329], '(201)': [1047, 1332], 'Our': [1050], 'goal': [1051], 'understand': [1054], 'how': [1055], 'like': [1057], 'GUS': [1058, 1270, 1287, 1302, 1382, 1585, 3199, 3458, 3682, 3795, 3910, 4038], 'XYL': [1060, 1589, 2937, 3124, 3461, 3665, 3851, 3871, 3903, 3914, 4051], 'become': [1063, 4041], 'so': [1064], 'specific.': [1065], 'X-ray': [1066], 'crystallography': [1067], 'essential': [1069], 'basis': [1074], 'specificity,': [1077], 'complexity': [1080], 'structures': [1083], 'necessitates': [1084], 'development': [1086], 'complementary': [1088], 'functional': [1089, 1113], 'approaches.': [1090], 'randomly': [1092, 1393, 2735], 'mutate': [1093], 'genes': [1094], 'or': [1095, 2024, 3637], 'parts': [1096], 'genes,': [1098], 'express': [1100], 'libraries': [1103], 'populations': [1105], 'microorganisms.': [1107], 'High': [1108], 'enables': [1111], 'evaluation': [1114], 'thousands': [1116], 'parallel.': [1121], 'Iterative': [1122], 'cycles': [1123, 2101, 2975], 'mutagenesis,': [1125, 1612], 'point': [1126, 1787], 'mutant': [1127], 'recombination,': [1128, 1425], 'lead': [1133], 'accumulation': [1136], 'beneficial': [1138], 'selected': [1141, 1778, 2537, 4024], '(13Rowe': [1143, 1893, 3480], 'L.A.': [1144, 1894, 2392, 3481, 3694], 'Geddie': [1145, 1895, 3482], 'M.L.': [1146, 1896, 2390, 3483, 3692], 'Alexander': [1147, 1897, 2393, 3484, 3695], 'O.B.': [1148, 1898, 2394, 3485, 3696], 'Matsumura': [1149, 1899, 3486], '2003;': [1154, 1687, 1904, 3491], '332:': [1155, 1905, 3492], '851-860Crossref': [1156, 1906, 3493], '(57)': [1159, 1909, 3496], 'Directed': [1162], 'alter': [1168], 'properties': [1169], 'including': [1172, 1884, 1933, 3659], 'substrate': [1173, 1497], '(reviewed': [1175], 'Refs.': [1177, 1613], '14Farinas': [1178], 'E.T.': [1179], 'Bulter': [1180], 'Arnold': [1182, 1567, 1616, 3002], 'F.H.': [1183, 1568, 1617, 3003], 'Biotechnol.': [1186, 1203, 1650, 1734, 3005, 3313], '12:': [1188], '545-551Crossref': [1189], '(231)': [1192, 1692], '15Zhao': [1195], 'Chockalingam': [1197], 'K.': [1198, 1566, 1615, 1662, 1666, 1680, 2832], 'Z.': [1200, 1668, 2999], '2002;': [1204, 1537, 1557, 1651, 1735], '13:': [1205], '104-110Crossref': [1206], '(151)': [1209], '16Schmidt-Dannert': [1212], 'Biochemistry.': [1214, 1756], '40:': [1216, 1260], '13125-13136Crossref': [1217], '(67)': [1220], '17Harris': [1223], 'J.L.': [1224], 'Craik': [1225], 'C.S.': [1226], 'Chem.': [1229, 1255, 1685], '1998;': [1231, 3006, 3611], '2:': [1232], '127-132Crossref': [1233], '(47)': [1236], '18Powell': [1239], 'K.A.': [1240], 'Ramer': [1241], 'Del': [1243], 'Cardayre': [1244], 'S.B.': [1245], 'Stemmer': [1246], 'W.P.': [1247, 1428, 2741, 3589], 'Tobin': [1248], 'M.B.': [1249], 'Longchamp': [1250], 'P.F.': [1251], 'Huisman': [1252], 'G.W.': [1253], 'Angew': [1254], 'Int.': [1256], 'Ed.': [1257], 'Engl.': [1258], '3948-3959Crossref': [1261], '(188)': [1264], 'good': [1273], 'model': [1274], 'system': [1275], 'study': [1278, 1601, 3479, 4126], 'evolution.': [1281, 1776, 1812], 'As': [1282, 3744], 'noted': [1283], 'above,': [1284, 2591], 'crystallized;': [1291], '50%': [1305], 'identical,': [1306], 'highly': [1308], 'sites': [1311, 2574], '(12Jain': [1312], '(gusA,': [1340], 'formerly': [1341], 'uidA)': [1342], 'more': [1344, 3951], 'amenable': [1345], 'evolution,': [1348], 'because': [1349, 3926, 4030], 'overexpressed': [1353], 'at': [1354, 2122, 2130, 2152, 2308, 2317, 2418, 2424, 2494, 2780, 3140, 3440, 3446, 3782, 3800, 3829, 3844, 3999], 'levels': [1356], 'enabling': [1360], 'detection': [1362], 'weak': [1364], 'β-xylosides': [1370], 'other': [1372], 'non-native': [1373], 'substrates.': [1374], 'expressed': [1389, 3799], 'mutated': [1394], 'gusA': [1395, 1950, 2036, 2647, 2851, 2891, 2963, 3018, 3129, 3155, 3187, 3193, 3472, 3759, 4148, 4249, 4306], 'alleles': [1396, 2648, 2964, 3130, 3167], 'lacZ-E.': [1399], 'strain.': [1401, 2531], 'Clones': [1402, 2412], 'LB': [1408], 'agar': [1409, 2431], 'plates': [1410, 2432, 3724], '5-bromo-4-chloro-3-indoyl-β-d-galactopyranoside': [1412], '(X-gal)': [1413], 'isolated.': [1415, 1454], 'After': [1416], 'additional': [1418, 2806, 3182], '(point': [1423], 'mutation': [1424, 1788], 'Ref.': [1426], '19Stemmer': [1427], '1994;': [1436, 2000, 2334, 2749, 2874], '91:': [1437, 2001, 2750, 2875], '10747-10751Crossref': [1438, 2751], '(1077)': [1441, 2754], 'screening,': [1445], '500-fold': [1448], 'increases': [1449, 3958, 4099], 'sequenced,': [1458], 'allele': [1462, 3156, 4149, 4250, 4307], 'contained': [1463, 2030, 4034], 'mutations.': [1465], 'dissected': [1467], 'effects': [1469], 'site-directed': [1474], 'found': [1477, 4086], 'four': [1479], 'changes': [1482, 1495], 'loop': [1487], '(T509A,': [1488], 'S557P,': [1489], 'N566S,': [1490], 'K568Q)': [1491], 'accounted': [1492], '(1Matsumura': [1499, 3526], 'Ellington': [1501, 3310, 3528, 4105], 'A.D.': [1502, 3311, 3529, 4106], '305:': [1507, 3534], '331-339Crossref': [1508, 3535], '(156)': [1511, 3538], 'extend': [1516], 'recently': [1518], 'developed': [1519], 'strategy': [1521, 1766], '(20Sio': [1522], 'C.F.': [1523], 'Riemens': [1524], 'A.M.': [1525], 'van': [1526], 'der': [1527], 'Laan': [1528], 'J.M.': [1529], 'Verhaert': [1530], 'R.M.': [1531], 'Quax': [1532], 'W.J.': [1533, 2356], 'Eur.': [1534], '269:': [1538], '4495-4504Crossref': [1539], '(44)': [1542], '21Lingen': [1545], 'Grotzinger': [1547], 'Kolter': [1549], 'Kula': [1551], 'M.R.': [1552, 2332], 'Pohl': [1553], 'Eng.': [1556], '585-593Crossref': [1559], '(61)': [1562], '22Miyazaki': [1565, 1614], '1999;': [1572, 1621, 3314], '49:': [1573, 1622], '716-720Crossref': [1574, 1623], '(176)': [1577, 1626], 'randomized': [1592, 1958], 'specificity-determining': [1593], 'our': [1599, 2384, 3564, 4082, 4131], 'previous': [1600, 3478, 4083], '(Ser557,': [1602], 'Asn566,': [1603], 'Lys568)': [1604], '(also': [1608], 'combinatorial': [1610], 'cassette': [1611], '23Coco': [1629], 'W.M.': [1630, 1990, 2864], 'Encell': [1631], 'L.P.': [1632], 'Levinson': [1633], 'W.E.': [1634], 'Crist': [1635], 'M.J.': [1636, 1682], 'Loomis': [1637], 'A.K.': [1638], 'Licato': [1639], 'L.L.': [1640], 'Arensdorf': [1641], 'J.J.': [1642, 1703], 'Sica': [1643], 'N.': [1644], 'Pienkos': [1645], 'P.T.': [1646], 'Monticello': [1647], '1246-1250Crossref': [1653], '(65)': [1656], '24DeSantis': [1659], 'Wong': [1661], 'Farwell': [1663], 'Chatman': [1665], 'Zhu': [1667], 'Tomlinson': [1669], 'Huang': [1671], 'Tan': [1673], 'X.': [1674], 'Bibbs': [1675], 'L.': [1676, 2997], 'Kretz': [1679], 'Burk': [1681], 'Am.': [1684], 'Soc.': [1686], '125:': [1688], '11476-11477Crossref': [1689], '25el': [1695], 'Hawrani': [1696], 'A.S.': [1697], 'Sessions': [1698], 'R.B.': [1699], 'Moreton': [1700], 'K.M.': [1701], 'Holbrook': [1702], '264:': [1708], '97-110Crossref': [1709], '(45)': [1712], '26Ness': [1715], 'J.E.': [1716], 'Kim': [1717], 'Gottman': [1719], 'Pak': [1721], 'Krebber': [1723], 'Borchert': [1725], 'T.V.': [1726], 'Govindarajan': [1727], 'Mundorff': [1729], 'Minshull': [1731], '1251-1255Crossref': [1737], '(165)': [1740], '27Graham': [1743], 'L.D.': [1744], 'Haggett': [1745], 'K.D.': [1746], 'Jennings': [1747], 'P.A.': [1748, 1755], 'Le': [1749], 'Brocque': [1750], 'D.S.': [1751], 'Whittaker': [1752], 'R.G.': [1753], 'Schober': [1754], '1993;': [1757, 3598], '32:': [1758], '6250-6258Crossref': [1759], 'effected': [1767], 'vast': [1768, 3906], 'improvements': [1769], 'single': [1772], 'further': [1781, 2120], 'random': [1784, 3936, 4088], '(Table': [1794], 'I).Table': [1795], 'ISummary': [1796], 'experiments': [1799, 3468, 3867], 'was': [1802, 1826, 1925, 1939, 2084, 2119, 2136, 2142, 2166, 2193, 2209, 2229, 2315, 2325, 2478, 2497, 2504, 2524, 2561, 2586, 2631, 2701, 2726, 2785, 2913, 2922, 3021, 3033, 3046, 3073, 3100, 3157, 3201, 3234, 3281, 3452, 3469, 3568, 3643, 3860, 3873, 3885, 3990, 4007, 4026, 4058, 4160, 4164, 4176, 4195, 4261, 4265, 4277, 4318, 4322, 4334], 'constructed': [1803, 3475], 'different': [1806, 3522, 3554], 'way': [1807], 'each': [1809, 1824, 1839, 2045, 2283, 2522, 2691, 3248, 3727, 3823, 3855, 3969, 4005, 4147, 4248, 4305], 'coefficient': [1816, 4192], '(improvement': [1817], 'fitness': [1819, 4073], 'ancestor)': [1822], 'population': [1825], 'determined': [1827, 1841, 3291, 3412, 4196], 'quantitative': [1829, 3952], 'cell': [1831, 2438, 3737], 'assays.': [1833], 'assayed': [1835], '16': [1836, 2422, 2620, 3780, 3848, 3966, 3997, 4014], 'replicates': [1837, 3967, 3981], 'clone,': [1840], 'those': [1845], 'immediate': [1852], 'ancestor,': [1853], 'divided': [1855], 'type.Round': [1863], 'evolutionDiversity': [1865], 'generationAverage': [1866], 'coefficient1Saturation': [1868], 'mutagenesis30-fold2Random': [1869], 'mutagenesis80-fold3DNA': [1870], '(catalytic': [1872], 'domain)90-fold4StEP/shuffling100-fold': [1873], 'Open': [1874, 3413], 'table': [1875, 3414], 'new': [1878, 3417], 'tab': [1879, 3418], 'Materials—Most': [1880], 'materials,': [1883], 'constitutive': [1886, 3471], 'His6-gusA': [1887], 'expression': [1888, 2037, 2845, 3473, 3760, 4039], 'vector,': [1889, 2038], 'described': [1892, 2350, 2550, 2590, 2643, 2820], 'oligonucleotides': [1913], 'synthesized': [1917], 'IDT': [1919], '(Coralville,': [1920], 'IA);': [1921], 'pNP-β-d-xylopyranoside': [1923, 2486, 3258], '(pNP-xyl)': [1924], 'Sigma-Aldrich.': [1927], 'GeneAmp': [1929], 'XL': [1930, 2049], 'polymerase': [1931, 1984, 2268, 2611], 'kit,': [1932], 'rTth': [1935], 'Vent': [1937, 1983], 'polymerases,': [1938], 'Applied': [1941, 3135, 3146], 'Biosystems': [1942, 3136], '(Foster': [1943], 'City,': [1944], 'CA).': [1945, 2223], 'Saturation': [1946, 3449], 'Mutagenesis—The': [1947], 'codons': [1948, 4151, 4252, 4309], 'encoding': [1949], '568': [1956], '(28Eisinger': [1963, 3574], 'D.P.': [1964, 3575], 'Trumpower': [1965, 3576], 'B.L.': [1966, 3577], 'BioTechniques.': [1967, 2333, 2833, 3578, 3610], '22:': [1969, 3580], '250-252,': [1970, 3581], '254Crossref': [1971, 3582], '(7)': [1974, 3585], 'mixture': [1979, 2083, 2700], 'Tth': [1981], '(29Cheng': [1985, 2859], 'Fockler': [1987, 2861], 'Barnes': [1989, 2863], 'Higuchi': [1991, 2865], '5695-5699Crossref': [2002, 2876], '(571)': [2005, 2879], 'back-to-back': [2009], '5′-phosphorylated': [2010], 'degenerate': [2011, 3631], 'primers:': [2012, 3183], '5′-GCGCAATATGCCTTGKNNGGTCGAAAATCGG-3′': [2013], '(gusA': [2014, 2018, 2669, 2884, 3025, 3037, 3041, 3053, 3190], 'S557X)': [2015], '5′-GTTGGCGGTNNKAAGNNKGGGATCTTCACTCGC-3′': [2017], 'N566X,K568X),': [2019], 'where': [2020], 'K': [2021, 2151, 3634], 'T': [2023, 3636], '50': [2031, 2148, 2187, 2288, 2673, 2769, 3268, 3815], 'ng': [2032, 2674], 'ancestral': [2035, 2202, 2530, 2844, 3464, 3758, 3918, 3984, 4220], '500': [2039, 2665], 'nm': [2040, 2043, 2282, 2420, 2496, 2666, 3243], 'primers,': [2041], '200': [2042, 2281, 2481], 'dNTP,': [2046, 2284, 2692], '1×': [2047], 'ABI': [2048], 'buffer': [2050, 3271], 'II,': [2051], '1.5': [2053], 'mm': [2054, 2289, 2294, 2297, 2485, 2679, 2684, 2687, 2690, 2770, 2775, 3269, 3812, 3816, 3994], 'magnesium': [2055], 'acetate.': [2056], 'overlaid': [2060, 2702], 'light': [2062], 'mineral': [2063, 2134, 2704], 'oil': [2064, 2135, 2705], 'heated': [2066], '80': [2068, 3763], '°C': [2069, 2094, 2104, 2109, 2124, 2154, 2319, 2426, 2621, 2712, 2718, 3846], 'thermal': [2072], 'cycler': [2073], 'hot': [2076], 'start.': [2077], '0.5': [2078], 'μl': [2079, 2188, 2473, 2482, 3740, 3809], 'Tth/Vent': [2082], 'added,': [2085], 'temperatures': [2088], 'immediately': [2090, 2786], 'raised': [2091], '94': [2093, 2103, 2711], '1': [2096, 2306, 3245, 3255, 3265], 'min,': [2097, 2322], 'followed': [2098, 3282], '30': [2100, 2714], '×': [2105, 2110, 3720, 3748], '15': [2106, 2156, 2683], 's,': [2107], '68': [2108], '5': [2111, 2270, 2296, 2500, 2694, 3242, 3739], 'min': [2112, 2114, 2127, 2157, 2779], '(1': [2113, 2603], 'per': [2115], 'kb).': [2116], 'reaction': [2118, 2141, 2585, 2803, 2818], 'incubated': [2121, 2143, 3828], '72': [2123, 2717], '10': [2126, 2293, 2321, 2774, 3262, 4021], 'stored': [2129], '4': [2131, 2254], '°C.': [2132, 2310, 2782, 3784, 4001], 'removed,': [2137, 3806], '0.5%': [2145], 'SDS': [2146], 'μg/ml': [2149], 'proteinase': [2150], '65': [2153, 2318], 'eliminate': [2159, 2200], 'thermostable': [2161], 'polymerases.': [2162], 'product': [2165, 2208, 2259, 2560, 2725, 2761, 2912, 3280, 3642, 4207], 'purified': [2167, 2257, 2326, 2727, 2954, 3217, 3249], 'Promega': [2170], 'Wizard': [2171, 2731], 'prep': [2173], 'kit': [2174], '(Madison,': [2175], 'WI)': [2176], 'manufacturer,': [2181], 'final': [2184], 'elution': [2185], 'water.': [2190], 'digested': [2194, 2563], 'restriction': [2196, 2565], 'DpnI': [2198, 2567], '(to': [2199, 2571], 'methylated': [2201], 'template),': [2203], 'desired': [2206], 'gel-purified': [2210], 'QiaQuick': [2213], 'spin': [2214], 'column': [2215], 'manufacturer': [2220], '(Qiagen,': [2221], 'Valencia,': [2222], 'concentration': [2225, 2598, 3233], 'eluted': [2227], 'estimated': [2230], 'via': [2231], 'agarose': [2232], 'gel': [2233], 'electrophoresis.': [2234], 'For': [2235], 'intramolecular': [2236], 'self-ligation': [2237], 'blunt-ended': [2239], 'DNA,': [2240, 2677], 'recommend': [2242], 'low': [2243], 'concentrations': [2244, 2249], 'ligase.': [2253], 'fmol': [2255], 'reacted': [2261, 2758, 3253, 3991], '2.5': [2263], 'units': [2264, 2272, 2695, 2764], 'Weiss': [2271], 'ligase': [2276, 2314, 2602], '20-μl': [2278], '100': [2285, 2974], 'μm': [2286, 2301, 3246, 3263], 'ATP,': [2287], 'Tris,': [2290, 3817], 'pH': [2291, 2681, 2772, 3818], '7.6,': [2292, 2773, 3819], 'MgCl2,': [2295, 2688, 2776], 'dithiothreitol,': [2298], '25': [2300, 2708], 'bovine': [2302, 2615], 'serum': [2303, 2616], 'albumin,': [2304], 'h': [2307, 2423, 3781, 3998], '23': [2309, 2625, 2781], 'heat-killed': [2316], 'butyl': [2328], 'alcohol': [2329], 'precipitation': [2330], '(30Thomas': [2331], '16:': [2335, 2365, 3007], '988-990PubMed': [2336], 'Freshly': [2339], 'prepared': [2340], 'InvαF′': [2343, 2630, 2921, 3108], 'cells': [2344, 3717, 3754, 3836], 'electroporation': [2348], 'Dower': [2352], 'et': [2353], 'al.': [2354], '(31Dower': [2355], 'Miller': [2357], 'J.F.': [2358], 'Ragsdale': [2359], 'C.W.': [2360], 'Nucleic': [2361], 'Acids': [2362], '1988;': [2364], '6127-6145Crossref': [2366], '(2157)': [2369], 'bacteria': [2374], 'titered,': [2376], 'distributed': [2377], '384-well': [2379, 3975], 'plates,': [2380], 'evaluated': [2382, 2505, 3680], 'microplate': [2387, 3689, 3712, 3864, 4214], 'assay': [2388, 3240, 3953], '(32Geddie': [2389, 3691], 'Rowe': [2391, 3621, 3693], 'Methods': [2397, 3699], 'Enzymol.': [2398, 3700], '388:': [2400, 3702], '134-145Crossref': [2401, 3703], '(11)': [2404, 3706], 'pNP-xyl': [2408, 3813, 3884, 3995], 'substrate.': [2411], 'associated': [2413, 2447, 2515, 3334, 3853, 3874], 'highest': [2416], 'absorbance': [2417, 2493], '405': [2419, 2495], 'after': [2421], '37': [2425, 3783, 3845, 4000], 'streaked': [2428, 2456], 'onto': [2429, 2457], 'LB-ampicillin': [2430, 2458, 3742], 'assay.': [2440], 'Whole': [2441, 2532], 'Cell': [2442], 'Activity': [2443], 'Assays': [2444], '(Secondary': [2445], 'Screens)—Microcultures': [2446], 'primary': [2453], 'screens': [2454], 'plates.': [2459], 'Individual': [2460], 'colonies': [2461], 'picked': [2463], 'grown': [2465], '96-well': [2470, 2489], 'microplate.': [2471], '20': [2472], 'saturated': [2476], 'culture': [2477], 'placed': [2479], '0.4': [2484], 'plate,': [2490, 3976], 'monitored': [2498], 'h.': [2501, 3849], 'Each': [2502, 3322, 3986], 'clone': [2503, 2523, 3970, 4006, 4068], 'three': [2507, 3056, 3340, 3511, 3559, 3939], 'independent': [2508, 3341], 'trials,': [2509], 'values': [2513, 3297, 3325, 4122], '(and': [2514, 3332], 'S.E.)': [2516, 3335], 'calculated.': [2518], 'normalized': [2525], 'Plasmid': [2533], 'PCR—The': [2534], 'first': [2540, 3500, 4140, 4156, 4257, 4314], 'purified,': [2545, 2562, 2914, 3101, 3644], 'PCR-amplified': [2548], 'above': [2551, 2821], '5′-GGGAAGCTTCTCATTGTTTGCCTCCCTGCTGCG-3′': [2553], '(3′-gusA3)': [2554], '5′-GGGAAGCTTGCGGCCGCACTCGAGCAC-3′': [2556], '(3′-pE-Tout3).': [2557], 'amplification': [2559, 3057], 'Hind': [2569], 'III': [2570, 3331], 'digest': [2572], 'underlined': [2575], 'primer': [2578, 3161], 'sequence),': [2579], 'gel-purified,': [2580], 'self-ligated.': [2582, 2917, 3104], 'ligation': [2584], 'except': [2592, 2972], 'lower': [2597], 'unit': [2604], 'instead': [2605], '5),': [2607], 'no': [2608, 2805], '(nor': [2612], 'dNTPs': [2613], 'albumin),': [2617], 'incubation': [2622], '(instead': [2623], '°C).': [2626], 'strain': [2629, 2920, 3107, 3651], 'library,': [2636, 2927, 3114], 'transformants': [2639, 2930, 3117], 'screened': [2641, 2932, 3119], 'above.': [2644], 'Shuffling—The': [2646], 'pooled': [2657, 2962], 'standard': [2662, 2816, 2969], 'primers': [2667, 2904, 3067, 3093, 3628], '5′-GGACTTTGCAAGTGGTGAATCCGCAC-3′': [2668, 3189], '720)': [2670], '3′-gusA3,': [2672], 'template': [2676], '60': [2678], 'Tris-HCl,': [2680, 2771], '8.5,': [2682], '(NH4)SO4,': [2685], '2.0': [2686], '0.2': [2689], 'Taq': [2697], 'polymerase.': [2698], 'cycled': [2707], 'times': [2709, 2981, 3560], 'between': [2710], 's': [2715], 'min.': [2721], 'according': [2728], 'procedure': [2733], 'recombined': [2736], '(19Stemmer': [2740], '0.1': [2763], 'DNase': [2766], 'I': [2767], 'extracted': [2787], 'phenol/chloroform': [2789], '(50:50)': [2790], 'ethanol': [2792], 'precipitated.': [2793], 'fragments': [2795, 2987], 'reassembled': [2797], '45': [2800], 'cycle': [2801], 'PCR-like': [2802], 'primers.': [2807], 'full-length': [2809, 2910], 'recombinant': [2810], 'subcloned': [2823], 'follows': [2828], '(33Temesgen': [2829], 'Eschrich': [2831], '21:': [2835], '828-830,': [2836], '832Crossref': [2837], '(5)': [2840], 'vector': [2846, 2895, 3072, 3081, 3474], '5′-CGGTTTGTGGTTAATCAGGAACTGTTCG-3′': [2883], '900out)': [2885], '3′-pETout3.': [2887, 2907, 3079], 'Finally,': [2888], 'shuffled': [2890], 'combined': [2897, 3059], '3′-gusA3': [2905], 'restriction-digested,': [2915, 3102], 'Staggered': [2939], 'Extension': [2940], 'Process': [2941], '(StEP)—Plasmids': [2942], 'derived': [2943], 'third': [2949], 'pooled.': [2956], 'Three': [2957], 'overlapping': [2958], 'regions': [2959], 'separately': [2967, 3252], 'reactions,': [2971], 'short': [2977], '(15': [2978], 's)': [2979], 'executed.': [2983], 'produces': [2985], 'incomplete': [2986], 'recombine': [2990], 'switching': [2992], 'templates': [2993], '(34Zhao': [2994], 'Giver': [2996], 'Shao': [2998], 'Affholter': [3000], 'J.A.': [3001], '258-261Crossref': [3008], '(585)': [3011], '5′-end': [3015], 'pool': [3020], '5′-TAATCACCATTCCCGGCGGGATAGTC-3′': [3024], '500out)': [3026], '5′-TCTAGATCTGGCACGACAGGTTTCCCGACTG-3′': [3028], '(rev137),': [3029], 'central': [3031], 'region': [3032], '5′-GCCATTTGAAGCCGATGTCACGCCG-3′': [3036], '360)': [3038], '5′-CCTGTAAGTGCGCTTGCTGAGTTTCC-3′': [3040], '1150out),': [3042], '3′-end': [3045, 3151, 4145, 4246, 4303], '5′-GGGAAGCTTATGTTTGCCTCCCTGCTGCGGTTTTTCA-3′': [3049], '(3′-gusA2)': [3050], '5′-CTGCTGCTGTCGGCTTTAACCTCTCT-3′': [3052], '1080).': [3054], 'rev137': [3068], '3′-gusA2.': [3070, 3096], '5′-AGCTGTTTCCTGTGTGAAATTGTTATCC-3′': [3076], '(rev0out)': [3077], 'insert': [3083], 'then': [3085], 'assembled': [3086], '3′-pETout3': [3094], 'improved': [3123], 'Sequencing—The': [3127], 'sequenced': [3132, 3158, 3175, 4165, 4266, 4323], 'Big': [3137], 'Dye': [3138], 'protocol': [3139], 'Center': [3142], 'Fundamental': [3144], 'Evolution': [3147], '(Emory': [3148], 'University).': [3149], 'every': [3153], '5′-GCTCAGCGGTGGCAGCAGCCAACTC-3′': [3162], '(GC': [3163], '3′pET).': [3164], 'Particularly': [3165], 'interesting': [3166], '(namely': [3168], '1.15,': [3169, 3211], '2.10,': [3170, 3212], '4.7,': [3171, 3213], '4.8)': [3173], 'entirety,': [3178], 'following': [3181], '5′-ATGCTTCCGGCTCGTATGTTGTGTGG-3′': [3184], '(rev': [3185], '80),': [3186], '360,': [3188], '720),': [3191], '1080.': [3194], 'Purification': [3196], 'Characterization—Each': [3198], 'fused': [3202], 'N-terminal': [3205], 'His6': [3206], 'tag,': [3207], '4.8': [3215], 'homogeneity': [3219], '(as': [3220], 'judged': [3221], 'SDS-PAGE': [3223], 'analysis)': [3224], 'nickel': [3226], 'chelate': [3227], 'affinity': [3228], 'chromatography.': [3229], 'total': [3231], 'quantified': [3235], 'Bradford': [3238], '(Bio-Rad).': [3241], 'ml': [3256], '(concentrations': [3259], 'ranging': [3260], 'mm)': [3266], 'Tris-HCl': [3270], '(pH': [3272], '7.6),': [3273], 'formation': [3276, 4208], 'pNP': [3279], 'spectrophotometer.': [3285, 3865], 'steady-state': [3287], 'kinetic': [3288], 'parameters': [3289, 3344], 'fitting': [3293], 'initial': [3295], 'velocity': [3296], 'Michaelis-Menten': [3300], 'equation': [3301], '(35Matsumura': [3302], 'Wallingford': [3304], 'J.B.': [3305], 'Surana': [3306], 'N.K.': [3307], 'Vize': [3308], 'P.D.': [3309], '17:': [3315], '696-701Crossref': [3316], '(62)': [3319], '(kcat,Km,': [3326], 'kcat/Km)': [3327], 'reported': [3328], 'Table': [3330, 4076], 'trials.Table': [3342], 'IIIKinetic': [3343], 'enzymesSubstrate': [3348], 'enzymepNP-β-d-xylopyranosidepNP-β-d-glucuronidekcatKmkcat/KmkcatKmkcat/Kms-1μ': [3349], 'ms-1m-1s-1μ': [3350], 'ms-1m-1WTaWT,': [3351], 'GUS0.0010': [3354], '±': [3355, 3357, 3359, 3361, 3363, 3365, 3367, 3369, 3371, 3373, 3375, 3377, 3379, 3381, 3383, 3385, 3387, 3389, 3392, 3394, 3396, 3398, 3400, 3402, 3404, 3406, 4224, 4226, 4228, 4230, 4232, 4234, 4236, 4238, 4240, 4242, 4244, 4296, 4298, 4300], '0.0001360': [3356], '602.9': [3358], '0.368': [3360], '6200': [3362], '10340,000': [3364], '20,0001.150.054': [3366], '0.003660': [3368], '5081': [3370], '30.039': [3372], '0.001210': [3374], '10190': [3376], '102.100.063': [3378], '0.005110': [3380], '10590': [3382], '300.042': [3384], '0.002180': [3386], '20240': [3388], '204.7NDbNot': [3390], 'determinedND240': [3391], '100.012': [3393], '0.001220': [3395], '1054': [3397], '24.8NDND190': [3399], '100.021': [3401], '0.004280': [3403], '9080': [3405], '10a': [3407], 'WT,': [3408], 'typeb': [3410], 'Not': [3411], 'Molecular': [3419], 'Modeling—We': [3420], 'visualized': [3421], 'crystal': [3422], 'structures,': [3423], 'made': [3424], 'sitedirected': [3425], 'silico': [3428], 'performed': [3430], 'energy': [3431], 'minimization': [3432], 'calculations': [3433], 'SYBYL': [3435], '(Tripos,': [3436], 'St.': [3437], 'Louis,': [3438], 'MO)': [3439], 'Biomolecular': [3442], 'Computing': [3443], 'Resource': [3444], '(BimCore)': [3445], 'Emory': [3447], 'University.': [3448], 'Mutagenesis—Our': [3450], 'objective': [3451], 'employed': [3501], 'randomize': [3507], 'residues:': [3512], 'encoded': [3542], 'twenty': [3544], '323': [3551], '(or': [3552], '32,768)': [3553], 'clones,': [3555], 'screen.': [3565], 'generated': [3569, 4094], '37Stemmer': [3588], 'Morris': [3590], 'S.K.': [3591], 'Kautzer': [3592], 'C.R.': [3593], 'Wilson': [3594], 'B.S.': [3595], 'Gene': [3596], '(Amst.).': [3597], '123:': [3599], '1-7Crossref': [3600], '(21)': [3603], '38Parikh': [3606], 'Guengerich': [3608], 'F.P.': [3609], '24:': [3612], '428-431Crossref': [3613], '(31)': [3616], '2L.': [3619], 'Matsumura,': [3624], 'submitted': [3625], 'manuscript.': [3626], 'NNK': [3632], '(where': [3633], 'G)': [3638], 'sequence.': [3639], 'self-ligated,': [3645], 'InvαF′.': [3652], 'K-12': [3655], 'derivatives,': [3658], 'InvαF′,': [3660], 'do': [3661], 'contain': [3663, 3935], 'endogenous': [3664], '(39Whitehead': [3667], 'T.R.': [3668], '35:': [3672], '282-286Crossref': [3673], '(16)': [3676], '∼10,000': [3681], 'semi-automated,': [3686], 'screen': [3690], 'dispensor': [3713], 'distribute': [3715], '77': [3719], '384': [3721, 3749], 'microtiter': [3723], 'such': [3725], '29,568': [3730, 3858], 'wells': [3731], 'received': [3732], 'one': [3736], 'medium.': [3743], 'control,': [3746], '3': [3747], 'microcultures': [3750, 3786, 3859, 3899], 'seeded': [3752], 'vector.': [3761], 'microplates': [3764, 3826], 'manually': [3766], 'sealed': [3767], 'autoclaved': [3769], 'silicone': [3770], 'seals': [3771, 3804], 'inverted': [3773], 'end-over-end': [3774], 'environmental': [3777], 'rotator': [3778], 'grew': [3787], 'under': [3790], 'conditions,': [3792], 'constitutively': [3798], 'levels.': [3802], '75': [3808], '1.0': [3811, 3993], 'added': [3821], 'well.': [3824], '45°': [3831], 'angle': [3832], '(so': [3833], 'settled': [3837], 'corner': [3840], 'wells)': [3843], 'measured': [3861], 'Reconstruction': [3866], 'showed': [3868], 'cells,': [3877], 'rather': [3878], 'supernatant,': [3881], 'suggesting': [3882], 'somehow': [3886], 'entering': [3887], 'cytoplasm.': [3889], 'Microsoft': [3891], 'Excel': [3892], 'macro': [3893], 'facilitated': [3894], 'identification': [3896], 'majority': [3907], 'less': [3913], 'controls': [3919], '3).': [3921], 'surprised': [3925], 'nearly': [3927], 'supposed': [3933], 'residues.': [3942], '24': [3944, 4023], 'reproducible': [3963], '(secondary': [3964], 'screen).': [3965], 'propagated': [3972], 'parallel': [3978], '48': [3980], 'clone.': [3985], 'cultures': [3989, 4200], 'calculated': [4008], 'averaging': [4010], 'ΔA405': [4012], 'replicates.': [4015], 'initially': [4018], 'reproducible,': [4028], 'perhaps': [4029], 'caused': [4037], 'genetically': [4042], 'unstable.': [4043], 'ten': [4045], 'discarded.': [4048], 'remaining': [4055], 'ancestor.': [4065], 'evinced': [4069], '70-fold': [4070], '(clone': [4074], '1.11,': [4075], 'II': [4077], 'Fig.': [4079], '4).': [4080], 'In': [4081, 4109], 'study,': [4084], '2–4-fold': [4098], '(40Matsumura': [4103], 'Braman': [4107], 'Vitro': [4110], 'Mutagenesis': [4111], 'Protocols.': [4112], '182.': [4113], 'Humana': [4114], 'Press,': [4115], 'Totowa,': [4116], 'NJ2001:': [4117], '259-267Google': [4118], 'current': [4125], 'validity': [4129], 'strategy.Table': [4134], 'IIAnalysis': [4135], 'evolutionVariant557566568Other': [4143], 'mutationsaThe': [4144], '(encoding': [4150, 4251, 4308], '440-604)': [4152, 4253, 4310], 'sequenced.': [4161, 4262, 4319], 'Clone': [4162, 4263, 4320], '1.15': [4163, 4264, 4321], 'entirety.': [4168, 4269, 4326], 'It': [4169, 4270, 4327], 'identical': [4171, 4272, 4329], 'variant': [4174, 4275, 4332], 'utilize': [4179, 4280, 4337], 'β-galactosides': [4180, 4281], '(1),': [4181, 4282], 'almost': [4184, 4285], 'certainly': [4185, 4286], 'result': [4187, 4288], 'cross-contaminationRelative': [4189], 'fitnessbThe': [4190], '(relative': [4193], 'fitness)': [4194], 'reacting': [4198], 'clonal': [4199], 'p-nitrophenyl-β-d-xylopyranoside,': [4202], 'measuring': [4203], 'rate': [4205], 'monitoring': [4210], 'A405': [4211], 'spectrophotometer': [4215], 'dividing': [4217], 'rateWTcWT,': [4221], 'typeSerAsnLys11.1LysAlaPhe23': [4223], '31.2GlnLysSerQ598stop3.3': [4225], '11.3ProSerVal30': [4227], '21.4TyrGluGly20': [4229], '51.5AspAsnPhe48': [4231], '41.7LeuCysMet33': [4233], '21.8AspAsnLeu46': [4235], '21.10SerSerLys21': [4237], '31.11CysSerLys72': [4239], '81.12CysSerLys30': [4241], '101.13ValCysMetD531E26': [4243], '21.15aThe': [4245], 'cross-contaminationProSerGlnS22N,': [4290], 'G81S,': [4291], 'K257E,': [4292], 'T509A,': [4293], 'Q598R,': [4294], 'stop604W25': [4295], '41.16IleAlaArgA580V11': [4297], '71.17ProCysPhe32': [4299], '5a': [4301], 'β': [4338]}, 'cited_by_api_url': 'https://api.openalex.org/works?filter=cites:W2113626439', 'counts_by_year': [{'year': 2023, 'cited_by_count': 3}, {'year': 2022, 'cited_by_count': 4}, {'year': 2021, 'cited_by_count': 3}, {'year': 2020, 'cited_by_count': 2}, {'year': 2018, 'cited_by_count': 2}, {'year': 2017, 'cited_by_count': 3}, {'year': 2016, 'cited_by_count': 3}, {'year': 2015, 'cited_by_count': 2}, {'year': 2014, 'cited_by_count': 1}, {'year': 2013, 'cited_by_count': 6}, {'year': 2012, 'cited_by_count': 2}], 'updated_date': '2024-12-08T09:15:34.613323', 'created_date': '2016-06-24'}