pET-33b质粒

基本信息

启动子： T7/lac

复制子： ColE1 ori，F1 ori

终止子： T7 terminator

质粒分类： 大肠杆菌载体；PET系列表达质粒

质粒大小： 5383bp

质粒标签： N-6×His, N-Thrombin，C-6×His

原核抗性： 卡那霉素Kan

克隆菌株： DH5α

培养条件： 37℃，有氧，LB

表达宿主： BL21(DE3)

诱导方式： IPTG或乳糖及其类似物

5'测序引物： T7：TAATACGACTCACTATAGGG

3'测序引物： T7-ter：TGCTAGTTATTGCTCAGCGG

备注： 含蛋白激酶PKA识别位点

质粒简介

The pET-33b(+) vector is derived from pET-28b(+) and carries a 15bp sequence encoding the protein kinase A (PKA) site RRASV, located between the thrombin cleavage and Nde I sites (1). Proteins expressed in pET-33b(+) can be easily purified by metal chelation chromatography (via either N- or C-terminal His•Tag® sequences) and efficiently labeled with 32P- or 33P-gATP and the catalytic subunit of cAMP-dependent protein kinase from heart muscle. Labeled proteins can be used as direct probes in protein-protein interaction studies. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/ expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer .