CAJ | 학술논문

目的构建人GMV/ARL-1真核表达载体,为进一步建立转ARL-1基因Hep-G2单克隆细胞系奠定基础.方法采用限制性内切酶技术,酶切真核骨架质粒pBK/CMV及另一质粒pBK/ARL-1(内含ARL-lcDNA全长),用TONA连结酶将目的基因连至骨架质粒上,酶切及序列分析的方法鉴定重组质粒.结果经Eco R I、Nco I、Uind Ⅲ酶切鉴定和序列分析,证明ARL-1基因已插入到pBK/CMV真核表达载体上.结论成功地构建了CHV/ARL-1真核表达载体.
목적구건인GMV/ARL-1진핵표체재체,위진일보건립전ARL-1기인Hep-G2단극륭세포계전정기출.방법채용한제성내절매기술,매절진핵골가질립pBK/CMV급령일질립pBK/ARL-1(내함ARL-lcDNA전장),용TONA련결매장목적기인련지골가질립상,매절급서렬분석적방법감정중조질립.결과경Eco R I、Nco I、Uind Ⅲ매절감정화서렬분석,증명ARL-1기인이삽입도pBK/CMV진핵표체재체상.결론성공지구건료CHV/ARL-1진핵표체재체.
Objective:To construct the eukaryotic expression vector CMV/ARL - 1 in order to prepare for establishing the Hep - G2 cell lines transfected with ARL - I. Methods: By endonuclease digestion, ARL - 1 gene from the plasmid pBK/ARL - 1 was cloned into the eukaryotic expression vector pBK/CMV to construct the eukaryotic expression vector CMV/ARL- I ,the recombinant plasmid was identified by endonuclease digestion and sequence analysis. Results:The ARL- I gene was identified to be inserted into the eukaryotic expression vector pBK/CMV by the digestion of Eco R Ⅰ , Nco Ⅰ , Hind Ⅲ and seqtence analysis. Conclusions: The eukaryotic expression vector CMV/ARL - 1 has been successfully constructed.