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大肠杆菌中噬菌体λ突变体的温度诱导

Temperature induction of bacteriophage lambda mutants in Escherichia coli.

作者信息

Chen B Y, Lin C S, Lim H C

机构信息

Department of Chemical and Biochemical Engineering, School of Engineering, University of California, Irvine 92717, USA.

出版信息

J Biotechnol. 1995 Jun 1;40(2):87-97. doi: 10.1016/0168-1656(95)00033-m.

Abstract

The paper presents temperature induction in Escherichia coli cells with phage lambda on the target-protein production and cell growth. Replicated lambda-DNA particles in the Q- and S- mutants remain naked for a longer time by preventing DNA packaging and cell lysis, and therefore the expression of the foreign genes is high. However, the parasitic infection of phage-lambda causes on significant losses of host cell viability in the induction phase. The temperature effects on cell growth and targeted-gene product formation were investigated. Gene amplification was found to be growth phase dependent for both Qam73 (Q mutation) and Sam100 (S mutation) mutants. Maximum induction occurs in the early exponential phase and under the optimal cell density. The total beta-galactosidase activity at this optimal induction condition increases roughly 8-10-fold with respect to that without thermal induction. To maximize the induction efficiency for the gene-product beta-galactosidase activity, several operating parameters were investigated. In this study, temperature induction is strongly dependent upon the population density of 'susceptible' cells at which time the temperature is shifted to 38-42 degrees C. This may be due to the 'threshold' population density to regulate the infection of lambda to hosts and control the productivity of target gene expression.

摘要

本文介绍了用噬菌体λ对大肠杆菌细胞进行温度诱导,以研究其对目标蛋白产生和细胞生长的影响。Q和S突变体中复制的λ-DNA颗粒通过阻止DNA包装和细胞裂解,在更长时间内保持裸露状态,因此外源基因的表达较高。然而,噬菌体λ的寄生感染在诱导阶段会导致宿主细胞活力显著损失。研究了温度对细胞生长和靶向基因产物形成的影响。发现基因扩增对Qam73(Q突变)和Sam100(S突变)突变体均呈生长阶段依赖性。最大诱导发生在指数生长期早期和最佳细胞密度条件下。在这种最佳诱导条件下,总β-半乳糖苷酶活性相对于无热诱导时大致增加8至10倍。为了使基因产物β-半乳糖苷酶活性的诱导效率最大化,研究了几个操作参数。在本研究中,温度诱导强烈依赖于“敏感”细胞的群体密度,此时温度转移到38-42摄氏度。这可能是由于“阈值”群体密度来调节λ对宿主的感染并控制目标基因表达的生产力。

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