含P_R启动子的原核高效表达载体的构建及应用  

Construction and Application of A High Level Expression Vector Containing P R Promoter.

作　　者：陈建军[1] 孙苗[1] 陈常庆[1] 卢芳[1] 王德宝[2] 

机构地区：[1]中国科学院上海生物工程研究中心,上海200233 [2]中国科学院上海生物化学研究所,上海200031

出　　处：《生物化学与生物物理进展》1999年第5期495-499,共5页Progress In Biochemistry and Biophysics

基　　金：国家"863"( 九.五) 计划资助

摘　　要：组建了一个仅含ＰＲ 启动子的原核高效表达载体ｐＲＣ， 它同时含有ｃⅠ调控基因、多酶切位点和两个强的转录终止序列． 现已成功地用于表达重组人肿瘤坏死因子α（ｈＴＮＦα）、重组人白细胞介素３ （ｈＩＬ３） 和抗溶菌酶（ＨＥＬ） 抗体Ｆｄ 基因， 表达量均占菌体总蛋白的３６％ 以上． 同时还研究了不同的宿主菌和原核增强子序列等因素对ＰＲ 启动子载体表达的影响． 此外， 还比较了分别以ＰＲ、ＰＬ 或ＰＲＰＬ 作为启动子时表达ｈＴＮＦα的情况，结果表明， 单用ＰＲ 或ＰＬ 启动子可获得与使用ＰＲＰＬA high level expression vector pRC has been constructed, which contains P R promoter, cIts857 gene, multiple cloning sites(MCS) and two strong transcription terminators. With this vector, hTNF α, hIL 3 and anti HEL McAb Fd genes have been expressed successfully, and their products each account for more than 36% of the total cell proteins. At the same time, it has been studied whether such factors as procaryon enhancer sequence and different strains affect the expression level of P R promoter vector . Furthermore, the inductive efficiency was compared among P R, P L and P RP L promoter, and the result has showed that P R or P L is as strong as P RP L.

关 键 词：启动子 载体构建 表达 增强子 

分 类 号：Q782[生物学—分子生物学]