Title: Construction of recombinant plasmid pcDNA3.1/hTSHRa
Abstract: Objective: To construct the recombinant eukaryotic expression vector pcDNA3.1/hTSHRa.Methods: Total thyroid RNA was prepared from normal human thyroid tissue.Then RNA was reversely transcripted and cDNA was subjected to PCR amplification.PCR product was cloned into pcDNA3.1 and then the recombinant vector was transformed into TOP10 E.coli.Constructed pcDNA3.1/hTSHRa was identified by PCR amplifying,restricting enzyme digestion analysis and DNA sequencing.Results: A 753 bp fragment encoding hTSHR ectodomain amino end was obtained by PCR amplification.PCR amplifying,HindⅢ restriction enzyme digestion,and DNA sequencing confirmed that pcDNA3.1/hTSHRa with the correct sequence and direction had been constructed successfully.Conclusion: hTSHRa with correct sequence and direction is cloned into the eukaryotic expression vector pcDNA3.1/hTSHRa,and it can be applied to protein expression in the future.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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