Construction of Human Myofibrillogenesis Regulator-1 Gene Eukaryotic Vectors and Its Expression in Neonatal Rat Cardiomyocytes
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    Abstract:

    AimTo construct the human myofibrillogenesis regulator-1(hMR-1) full length eukaryotic expression plasmid and measure its expression in HEK293T cell lines and Sprague-Dawley neonatal rat cardiomyocytes.MethodsThe whole hMR-1 coding gene was cloned from NCBI GenBank database (AF417001), recombined with plasmid pcDNA3.1/Myc-His(-)B and transformed into Escherichia coli XL1-Blue.Positive clone was selected and sequenced by T7 primers.The recombined plasmid pcDNA3.1/Myc-His(-)B -hMR-1 was transfected by Lipofectamin2000 to cells, reverse transcription polymerase chain reaction (RT-PCR) and Western Blotting was employed for measuring the expression of hMR-1.ResultsPlasmid pcDNA3.1/Myc-His(-)B-hMR-1 was verified by sequencing that the target gene was correct, without any mutation; the expression level at mRNA and protein were both increased significantly after hMR-1 transfection.ConclusionA full-length hMR-1 coding gene eukaryotic vector was successfully constructed, based on which, a convenient and effective transient transfection method in neonatal rat cardiomyocytes was established as well.

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WANG Xiao-Reng, LIU Xiu-Hua, LI Ting,,HAN Wen-Ling. Construction of Human Myofibrillogenesis Regulator-1 Gene Eukaryotic Vectors and Its Expression in Neonatal Rat Cardiomyocytes[J]. Editorial Office of Chinese Journal of Arteriosclerosis,2010,18(2):91-94.

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  • Received:January 18,2010
  • Revised:January 18,2010
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