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来自新疆嗜盐红菌的古紫质基因在大肠杆菌中的基因克隆与功能表达

Genetic cloning and functional expression in Escherichia coli of an archaerhodopsin gene from Halorubrum xinjiangense.

作者信息

Feng Jie, Liu Hong-Can, Chu Jin-Fang, Zhou Pei-Jin, Tang Ji-An, Liu Shuang-Jiang

机构信息

State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Zhong-Guan-Cun, Haidian, Beijing, 100080, People's Republic of China.

出版信息

Extremophiles. 2006 Feb;10(1):29-33. doi: 10.1007/s00792-005-0468-x. Epub 2005 Aug 2.

Abstract

Pairs of PCR primers that targeted the archae/bacteriorhodopsin gene were used to clone the archaerhodopsin (aR) gene of Halorubrum xinjiangense strain BD-1(T), and this gene was sequenced and functionally expressed in Escherichia coli. Recombinant E. coli cells harboring the plasmid carrying this gene became slightly purple or blue depending on whether they were supplemented with all- trans retinal or 3,4-dihydroretinal, respectively, during induction with IPTG. The purple and blue membranes from the recombinant E. coli showed maximal absorption at 555 and 588 nm, respectively, which are different from maximal absorption at 568 nm of the wild-type purple membrane. Purple membranes from the recombinant E. coli and from strain BD-1(T) were investigated in parallel. The E. coli purple membrane was fabricated into films and photoelectric responses were observed that depended on the light-on and light-off stimuli.

摘要

使用靶向古菌/细菌视紫红质基因的PCR引物对,克隆了新疆嗜盐红菌BD-1(T)菌株的古菌视紫红质(aR)基因,并对该基因进行测序,然后在大肠杆菌中进行功能表达。携带该基因质粒的重组大肠杆菌细胞,根据在异丙基-β-D-硫代半乳糖苷(IPTG)诱导过程中分别添加全反式视黄醛或3,4-二氢视黄醛,会变成浅紫色或蓝色。重组大肠杆菌的紫色膜和蓝色膜分别在555和588nm处显示出最大吸收峰,这与野生型紫色膜在568nm处的最大吸收峰不同。对重组大肠杆菌和BD-1(T)菌株的紫色膜进行了平行研究。将大肠杆菌紫色膜制成薄膜,并观察到其光电响应取决于光开和光关刺激。

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