人乳铁蛋白基因的克隆及序列分析与重组逆转录病毒质粒载体PLNCX2-hLF的构建  

Cloning and sequence analysis of human lactoferrin and construction of the recombinant retroviral vector of PLNCX2-hLF

作　　者：陈绍威[1] 字向东[1] 梁冠男[1] 尹荣华[1] 张大伟[1] 

机构地区：[1]西南民族大学生命科学与技术学院,四川成都610041

出　　处：《西南民族大学学报（自然科学版）》2008年第4期700-703,共4页Journal of Southwest Minzu University(Natural Science Edition)

基　　金：四川省学术和技术带头人培养资金助项目

摘　　要：从正常人外周血中性粒白细胞中提取总RNA,用RT-PCR的方法扩增人乳铁蛋白(hLF)cDNA,将其克隆到pMD18-T Simple载体上并测序.用限制性内切酶XhoⅠ和HindⅢ将目的片段与逆转录病毒载体PLNCX2进行双酶切,并用T4连接酶进行连接;然后转化到top10菌中,再次进行DNA序列测定.前后两次序列测定结果一致,没有突变,全长为2,136bp,说明所克隆的为hLF基因的cDNA序列;与GeneBank中登录的序列相比,同源性达99%以上.White blood cells are isolated from normal human peripheral bloods and total RNA is extracted. The cDNA are amplified by RT-PCR and then cloned into pMD18-T simple vector. After being sequenced, digest the fragment and retroviral vector by restriction enzymes Xho Ⅰ and Hind Ⅲ. The conjugate is transferred into E, coli cell line Top 10 and sequenced again Both sequencings show that the cloned gene sequence hLF consistent, no mutation, 99 % homology with other sequences in GeneBank.

关 键 词：中性粒白细胞 人乳铁蛋白 cDNA克隆 逆转录病毒载体 DNA序列分析 

分 类 号：Q81[生物学—生物工程]