Title: Establishment and Expression Analysis of miR-1 Adenovirus Vector
Abstract: Objective To construct and identify heart-enriched microRNAs adenovirus vector containing miR-1 precursor gene.Methods The primers of miR-1 precursor was designed and used for PCR amplification.The products were then cloned into adenovirus shuttle plasmid pAdTrack and linearized by enzyme Pme I;the resultant plasmid was co-transfected into E coli BJ51 83cells with adenovirus backbone plasmid pAdEasy-1 for homologous recombination.Then the recombinant plasmid was identified,1 inearized and packaged with QBI-293 Acells to amplify the recombinant adenovirus Ad-miR-1 ,which was then used to infect cardiomyocytes.Real-time quantitative PCR was used to detect the expression of miR-1 .Results miR-1 recombinant plasmid was constructed successfully as confirmed by sequencing and enzyme digestion.Real-time quantitative PCR confirmed that adenovirus Ad-miR-1 can significantly enhance the intracellular expression of miR-1 in cardiomyocytes.Conclusion The adenovirus that can express miR-1 has been successfully constructed and transfection with it into cardiomyocytes can elevate the expression level of miR-1.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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