人血清白蛋白基因的克隆及其真核表达载体构建.pdfVIP

福建畜牧兽医 第35卷 第5期 2013年 1 人血清白蛋白基因的克隆及其真核 表达载体构建 洪志勇 李玉芳1 1，2 张兴峰 林秀娇 李鸿翔 庄益芬 张文昌1 1 1 1 1* （1.福建农林大学动物科学学院 福州 350002；2.黄淮学院 河南驻马店 463000） 摘 要 利用RT-PCR扩增并克隆了含人血清白蛋白序列长约 1.8kb的片段，酶切鉴定并进行序列分析，测序结果与GenBank 中登录的人血清白蛋白cDNA序列的同源性为97%；以鹌鹑卵清蛋白5’调控区POV来启动HAS的表达。用限制性内切酶kpn 玉和Hind芋双酶切质粒pOV和pHSA，然后用T4连接酶定向连接成重组质粒pOV-pHSA，经酶切鉴定，两者成功融合。 关键词 人血清白蛋白 鹌鹑 卵管特异性表达载体 文献标识码：A 文章编号：1003-4331（2013）05-0001-05 Genecloningof humanserumalbuminandthe constructionofeukaryoticexpressionvector 1 1，2 1 HongZhiyong LiYufang ZhangXingfeng 1 1 1 1* LinXiujiao LiHongxiang ZhuangYifen ZhangWenchang （1.CollegeofAnimalScience，FujianAgricultureand ForestryUniversity，Fuzhou 350002； 2.HuanghuaiUniversity，Zhumadian，Henan 463000） Abstract About 1.8 kb sequence of the fragment of human serum albumin containing were successfully amplified by RT-PCR.The purpose of the initia gene were identified with the help of electrophoresis. The DNA fragment was cloned into vector pGEM-T and transformed into E.coli DH5a host bacteria. Xba 玉endonuclease digestion and sequencingfrom selected positive cloned bacteria. The fragment was sequenced，and it was compared with serum albumin cDNA gene of humanin GenBank.The results indicated the homology were 97%.Using 5’-flanking of ovalbumin in quail for the control sequence to activate the expression of HAS.With restrictive interior contact enzyme kpn 玉with Hind芋cuts material particle pOV and pHSA，then connects reorganization material particleforpOV-pHSAwiththeT4 ligase.Thefusionvectorwas sequencedbyenzymedigestion. Keywords human serumalb