pBI121载体酶切位点添加和拟南芥Na+H+逆向转运蛋白基因表达载体.pdfVIP

分子植物育种，2006 年，第4 卷，第6 期，第811-818 页 Molecular Plant Breeding, 2006, Vol.4, No.6, 811-818 研究报告 Research Report + + pBI121 载体酶切位点添加及拟南芥Na /H 逆向转运蛋白基因表达载体 的构建 1,2 1,2* 1,2 3 张俊莲 王蒂 张金文 陈正华 1 甘肃农业大学农学院, 兰州, 730070; 2 甘肃省作物遗传改良与种质创新重点实验室, 兰州, 730070; 3 甘肃亚盛集团博士后科研工作站北京分站, 北京, 100101 * 通讯作者, wangd@ 摘 要 对pBI121 载体GUS基因后的终止子序列 ( Ⅰ- R Ⅰ) 通过PCR 扩增法进行了酶切位点的添 Sac Eco 加，即在 Ⅰ后添加了 Ⅰ、在 R Ⅰ前添加了 Ⅰ。序列分析发现，两个酶切位点的添加是成功的， Sac Xho Eco Kpn 但扩增的终止子序列与原序列的同源性间有差异，即有4 个碱基发生了变异，特别是第 17 个碱基位点出现 了缺失。利用绿色荧光蛋白(GFP)对添加酶切位点载体进行了终止子活性的检测，结果发现，它与携带GFP 的pBI121 载体一样， 基因能正常表达，说明pBI121 载体终止子序列酶切位点的添加是成功的。利用改 GFP 造后的载体(pBI121-GZ)构建了CaMV35S 启动子驱动AtNHX1 基因的植物表达载体。 关键词 pBI121, 终止子, 限制性酶, 绿色荧光蛋白, 植物表达载体 + + Modification of pBI121 Vector and Expression Vector Construction Na /H Antiporter of Arabidopsisthaliana Zhang Junlian1,2 Wang Di1,2 Zhang Jinwen1,2 Chen Zhenghua3 1 Agronomic College of Gansu Agricultural University, Lanzhou, 730070; 2 Gansu Key Laboratory of Crop Genetic Germplasm Enhancement , Lanzhou, 730070; 3 Postdoctor Workstation of Yasheng Industrial Ltd., Beijing, 100101 * Corresponding author, wangd@ The two exonuclease ( Ⅰand Ⅰ) were added to NOS-ter ( Ⅰ- R Ⅰ) behind of Abstract Xho Kpn Sac Eco GUS pBI121 vector by PCR ．The sequence analysis showed that it was a success for adding to exonuclease, but NOS-ter sequence by PCR differed from original sequence, that meant NOS-ter sequence by PCR had four muta- tion sites, specially 17 site deletion ．However, the termination signal o