title: Genetic cloning and functional expression in Escherichia coli of an archaerhodopsin gene from Halorubrum xinjiangense.
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.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Zhong-Guan-Cun, Haidian, Beijing, 100080, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Feng
authors.forename: Hong-Can
authors.initials: HC
authors.lastname: Liu
authors.forename: Jin-Fang
authors.initials: JF
authors.lastname: Chu
authors.forename: Pei-Jin
authors.initials: PJ
authors.lastname: Zhou
authors.forename: Ji-An
authors.initials: JA
authors.lastname: Tang
authors.forename: Shuang-Jiang
authors.initials: SJ
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D019641:DNA, Archaeal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D019766:Genes, Archaeal; D006216:Halobacteriaceae / Q000235:genetics*; D008969:Molecular Sequence Data; D010777:Photochemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry; D017494:Proton Pumps
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D011994:Recombinant Proteins; C055476:archaerhodopsin protein, Archaea; D017494:Proton Pumps