C－myc基因外显子1导入并稳定高效表达的细胞株的建立

将ｃ－ｍｙｃ外显子１的ＤＮＡ片段，插入带有ｎｅｏ基因的真核细胞表达载体ｐＲＣ／ＣＭＶ，运用电穿孔基因导入法将此构建好的重组质粒导入Ｈｅｌａ细胞。在抗Ｇｅｎｅｔｉｃｉｎ的Ｈｅｌ３细胞抽提物中经免疫印迹技术检测，发现一条分子量约为３２ＫＤ的蛋白带，从而实现了ｃ－ｍｙｃ外显子１在真核细胞中高效、稳定地表达，为进一步研究ｃ－ｍｙｃ外显子１的生理功能打下基础。

INTRODUCTION AND EXPRESSION OF FIRST EXON OF HUMAN C-MYC PROTO ONCOGENE IN EUKARYOTIC CELL LINE

uman c-myc proto-oncogene has a coding capactity for a polypeptide of 188 residueswithin the first exon. The product of this exon may play a role in the regulation of cellgrowth and differentiation.In order to investigate the mechanism of this protein,a recombi-nant plamid was constructed for expression of first exon of c-myc gene. The DNA fragmentof first exon of c-myc gene was inserted into eukaryotic expression vector,pRC/CMV,whichbears aminoglycoside phosphotransferase gene(neo gene),and then,the well constructedrecombinant plasmid,pRC/CMV-exl, was introduced into Hela cell line by electroporation.The expression of first exon of c-myc gene was driven by CMV1 promoter. The colonies thatresisted G418 were visible after 10 days of transformation.A protein band of 32KD was de-tected by Western blot analysis in most colonies.