【摘要】 本文介绍了一种新型真核表达载体的构建过程。原核质粒载体ｐＳＰ７２经亚克隆去除部分单酶切点后，在其上定向插入真核选择标记ｇｐｔ基因片段，获得中间载体ｐＳＰ７２Ａ－ｇｐｔ。将来自表达载体ｐＭＡＭ－ｎｅｏ质粒上约２．４ｋｂ的真核表达启动、剪接与终止区（包括ＲＳＶ和ＭＭＴＶ－ＬＴＲ）克隆到ｐＳＰ７２Ａ－ｇｐｔ上，获得了一种新型真核表达载体ｐＳＭＧ（ｐＳＰ７２Ａ－ｇｐｔ－ＭＭＴＶ）。该载体在真核细胞中可受激素诱导而高效表达插入到其多克隆位点上的外源基因，并能与ｎｅｏ（Ｇ４１８）、ＤＨＦＲ、ＡＤＡ等为选择标记的真核表达载体配合使用，将两种以上的外源基因导入同一真核细胞，为研究多基因间的相互作用提供了一种良好手段。更多还原

【Abstract】 In this paper,the construction process of a new kind of mammalian expression vector is described. Prokaryotic amplification vector pSP72A, Which was generated by deleting some single restriction endonuclease sites from pSP72 plasmid,was recombined with eukaryotic selecting marker gpt gene fragment to get the intermediate vector pSP72A-gpt.The final vector pSMG (pSP72A-gpt-MMTV)was then constructed by inserting 2.4kb initation ,splicing and termination region(derived from pMAMneo,including RSV and MMTV)for eukaryotic gene expressions into pSP72A-gpt. This vector can express exogenous genes cloned in its MCS (multiple cloning sites) in many kind of cultured cells and can be induced at high level expression by administration of glucocorticoid. Combining pSMG with other mammalian expression vectors containing different selecting markers [Such as aminoglycoside phosphotransferase (neo or G418), dihydrofolate reductase (DHFR),adenosine deaminase (ADA) etc. ],two or more kinds of exogenous genes can be easily introduced into the same cell line and this will provide an useful tool for studying multiple genes interaction in eukaryotic cells.更多还原