CAJ | 학술논문

MEF2C基因能够控制肌细胞分化过程中的基因转录,特别在骨骼肌、心肌和平滑肌中介导细胞的分化。本研究将猪 MEF2C 基因化学合成后经 Bam HI 和 Asc I 双酶切后克隆到慢病毒表达载体 pLen-ti6.3_MCS_IRES2-EGFP中，获得的重组质粒用限制性内切酶酶切分析和测序鉴定；利用慢病毒阴性对照侵染靶细胞C2C12来确定最佳MOI值以及靶细胞最适抗生素Blasticidin剂量。结果显示猪MEF2C基因成功克隆到慢病毒表达载体中；该重组质粒侵染靶细胞C2C12的最佳MOI值为300；靶细胞抗生素的筛选剂量为4μg/mL，维持剂量为3μg/mL上述试验为建立MEF2C稳定过表达细胞系提供了前期工作基础。
MEF2C기인능구공제기세포분화과정중적기인전록,특별재골격기、심기화평활기중개도세포적분화。본연구장저 MEF2C 기인화학합성후경 Bam HI 화 Asc I 쌍매절후극륭도만병독표체재체 pLen-ti6.3_MCS_IRES2-EGFP중，획득적중조질립용한제성내절매매절분석화측서감정；이용만병독음성대조침염파세포C2C12래학정최가MOI치이급파세포최괄항생소Blasticidin제량。결과현시저MEF2C기인성공극륭도만병독표체재체중；해중조질립침염파세포C2C12적최가MOI치위300；파세포항생소적사선제량위4μg/mL，유지제량위3μg/mL상술시험위건립MEF2C은정과표체세포계제공료전기공작기출。
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.