TIGHTLY REGULATED TAC PROMOTER VECTORS USEFUL FOR THE EXPRESSION OF UNFUSED AND FUSED PROTEINS IN ESCHERICHIA-COLI
TIGHTLY REGULATED TAC PROMOTER VECTORS USEFUL FOR THE EXPRESSION OF UNFUSED AND FUSED PROTEINS IN ESCHERICHIA-COLI
复制标题
DOI：
10.1016/0378-1119(88)90440-4
复制
发表时间：
1988-09-30
期刊：
影响因子：
3.5
通讯作者：
ABEL, KJ
中科院分区：
文献类型：
--
作者：
AMANN, E;OCHS, B;ABEL, KJ
构建了一系列新的质粒表达载体（pTrc系列），用于调控大肠杆菌基因的表达。基于pKK233-2 [Amann and Brosius， Gene 40(1985) 183-190]，载体携带强杂交trp/lac启动子、lacZ核糖体结合位点（RBS）、pUC18多克隆位点和rrnB转录终止子。在合成的寡脱氧核苷酸的帮助下，在三个阅读框的NcoI位点后面插入了多个克隆位点。因此，载体对于表达真实的非融合形式的蛋白质（通过使用NcoI位点）和表达融合蛋白（通过选择正确翻译框架中的任何克隆位点）同样有用。为了确保在任何寄主菌株的构建和生长过程中完全抑制杂交trp/lac启动子，在一些载体上添加了lac抑制基因的lacIq等位基因。介绍了新载体的完整核苷酸序列和异源基因（人凝血因子XIIIa和人胎盘抗凝蛋白PP4）的表达实例。
A series of new plasmid expression vectors (the pTrc series) has been constructed for the regulated expression of genes in Escherichia coli. Based on pKK233-2 [Amann and Brosius, Gene 40 (1985) 183-190], the vectors carry a strong hybrid trp/lac promoter, the lacZ ribosome-binding site (RBS), the multiple cloning site of pUC18 and the rrnB transcription terminators. With the aid of synthetic oligodeoxynucleotides, the multiple cloning site has been inserted behind an NcoI site in three reading frames. Thus, the vectors are equally useful for the expression of proteins in their authentic, non-fused form (by using the NcoI site) and for the expression of fusion proteins) by choosing any of the cloning sites in the correct translational frame). To ensure complete repression of the hybrid trp/lac promoter during construction and growth in any host strain, the lacIq allele of the lac repressor gene was added to some of the vectors. The complete vector nucleotide sequence and examples of heterologous gene expression (human coagulation factor XIIIa and human placental anticoagulant protein PP4) with the new vectors are presented.