Detection and expression of a gene encoding a new bacteriorhodopsin from an extreme halophile strain HT (JCM 9743) which does not possess bacteriorhodopsin activity |
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Authors: | M Kamekura Yukio Seno Hiroaki Tomioka |
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Institution: | (1) Noda Institute for Scientific Research, 399 Noda, Noda-shi, Chiba-ken 278, Japan Tel. +81-471-23-5573; Fax +81-471-23-5959 e-mail: mkamekur@supernig.nig.ac.jp, JP;(2) Department of Chemistry, Faculty of Education, Saitama University, Urawa-shi, Saitama-ken 338, Japan, JP |
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Abstract: | Membrane vesicles prepared from an extreme halophile strain, HT (JCM 9743), showed no bacteriorhodopsin activity. However,
a DNA fragment, amplified by polymerase chain reaction (PCR), appeared to encode the C to G helices of a bacteriorhodopsin(bR)-like
protein. With the PCR product as a probe, the gene coding for a novel bacteriorhodopsin was cloned from the genomic DNA of
the strain HT. The open reading frame of the gene was ligated with the promoter region of the bop gene of Halobacterium salinarum bR, and expressed in a bR-deficient host strain, L33, using the plasmid vector pXLNov-R. The purplish membrane fraction purified
from cells of a transformant exhibited a cyclic photoreaction characteristic of bacteriorhodopsin.
Received: August 12, 1997 / Accepted: October 20, 1997 |
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Keywords: | Bacteriorhodopsin Extreme halophile Halobacterium salinarum Vector pXLNov-r Photocycle |
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