摘要
以 ZG1(+ )、ZG1(-)、ZG2 (+ )、ZG2 (— )、ZG3 (+ )、ZG3 (-)寡聚核苷酸片段为材料 ,合成了含有 GCN4亮氨酸拉链蛋白基区结合 DNA必需的 3 5个氨基酸的折叠片段 ,将此片段分别克隆到 p ET3 b质粒和 p BL Z质粒中 ,酶切后用 2 %琼脂糖电泳检测证明两者克隆成功 .将这 2种重组质粒分别转化到 E.coli DH5α中 ,发现 p ET3 b重组体在 E.coli DH5α中表达成功 ,而 p BL Z质粒重组体在 E.coli DH5α中不能成功表达 ;从转化子中分离得到重组质粒 p ET3 b。
Oligonucleotides ZG1(+),ZG1(-),ZG2(+),ZG2(—),ZG3(+ )and ZG3(-)were used to synthesize the leucine-zipper protein domain with C G 4 containing the 35 kinds of amino acids necessary for DNA binding. The synt hesize d d omain was cloned respectively to plasmid pET3b and pBLZ, then transformed into E.coli DH5α. They are found to be successfully cloned with 2 % agar ose gel electrophoresis by digested reconstructed plasmids with BamHⅠand NdeⅠ. The reconstructed pET3b could successfully express in E.coli DH5α,but the reconstructed pBLZ failed to express in E.coli DH5α.It is iden tified that the domain inserted to pET3b is the synthesized genes by restricted enzyme and sequence analysis.
出处
《中南民族大学学报（自然科学版）》
CAS
2003年第4期9-11,共3页
Journal of South-Central University for Nationalities：Natural Science Edition
基金
国家留学基金委资助项目 (99842 13 5)
关键词
亮氨酸拉链结构
基因合成
克隆
载体
表达
leucine zipper protein
gene synthesis
cloning
vector
expression
