Construction of the lentiviral expression vector of porcine MEF2C gene and optimization of transfection conditions in C2C12 cells
Zhu Hongyan
Yang Huixin
Tian Yumin
Su Yuhong
Abstract：MEF2C gene can control the genetic transcription during the differentiation of myo-cyte, especially can mediate cell differentiation in skeletal, cardiac and smooth muscle. The che-mosynthesized MEF2C gene was digested with the double restriction enzymes Bam HI and Asc I. The ob-tained MEF2C gene was inserted into the lentiviral expression vector pLenti6.3_MCS_IRES2-EGFP. The recombinant plasmid was identified by restriction enzymes digestion analysis and DNA sequencing. We used a negative control of the lentivirus to infect the target C2C12 cells in order to deter-mine the best MOI value and the optimal dose of antibiotics of the target cells. The results showed that the MEF2C gene expression vector was constructed successfully. The optimal MOI value of target cells was 300. The Blasticidin screening dose of target cells was 4 μg/mL and the sus-taining dose was 3 μg/mL.The above-mentioned researches provided a foundation for the further study to establish stable overexpressed cell lines.
Keywords：SwineMEF2C geneLentiviral expression vectorC2C12 cells
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 1-4 )