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[基于噬菌体λDNA中温度调控的P'R启动子的表达载体的构建及特性]

[Construction and properties of the expression vector based on the temperature-regulated P'R promoter in phage lambda DNA].

作者信息

Petrenko L A, Gileva I P, Kravchenko V V

出版信息

Bioorg Khim. 1987 Nov;13(11):1561-9.

PMID:2964824
Abstract

Plasmid expression vector using the temperature-regulated promoter P'R of bacteriophage lambda is described. The vector carries a combination of two regions of lambda cI857indgenome, that contain: 1) gene cI and promoter PR, and 2) gene Q and promoter P'R. Transcription or gene Q is initiated at promoter PR, which is controlled by cI857 repressor. The Q gene product acts as a positive regulator of RNA synthesis from P'R. At 37 degrees C, sufficient amounts of protein Q are synthesised to initiate the expression of the cloned gene from P'R. Inactivation of Q gene (by elimination of a single NcoI site) results in the loss of P'R expression activity in the vector. E. coli beta-galactosidase gene (lacZ) and human leukocyte interferon alpha 2 gene (ifn alpha 2) were cloned into a single EcoRI cleavage site under the control of P'R. These constructs express high levels of beta-galactosidase and interferon alpha 2 in E. coli at 37 degrees C.

摘要

描述了使用噬菌体λ的温度调节启动子P'R的质粒表达载体。该载体携带λcI857基因组两个区域的组合,其中包含:1)基因cI和启动子PR,以及2)基因Q和启动子P'R。基因Q的转录在启动子PR处起始,该启动子受cI857阻遏物控制。Q基因产物作为来自P'R的RNA合成的正调控因子。在37℃时,合成足够量的蛋白质Q以起始来自P'R的克隆基因的表达。Q基因的失活(通过消除单个NcoI位点)导致载体中P'R表达活性的丧失。大肠杆菌β-半乳糖苷酶基因(lacZ)和人白细胞干扰素α2基因(ifnα2)在P'R的控制下克隆到单个EcoRI切割位点。这些构建体在37℃时在大肠杆菌中高水平表达β-半乳糖苷酶和干扰素α2。

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