Abstract:
:A DNA region carrying lysS, the gene encoding the lysyl-tRNA synthetase, was cloned from the extreme thermophile prokaryote Thermus thermophilus VK-1 and sequenced. The analysis indicated an open reading frame encoding a protein of 492 amino acids. This putative protein has significant homologies to previously sequenced lysyl-tRNA synthetases and displays the three motifs characteristic of class II aminoacyl-tRNA synthetases. The T. thermophilus lysS gene was overexpressed in Escherichia coli by placing it downstream of the E. coli beta-galactosidase gene promoter on plasmid pBluescript and by changing the ribosome-binding site. The overproduced protein was purified by heat treatment of the crude extract followed by a single anion-exchange chromatography step. The protein obtained is remarkably thermostable, retaining nearly 60% of its initial tRNA aminoacylation activity after 5 h of incubation at 93 degrees C. Finally, lethal disruption of the lysRS genes of E. coli could not be compensated for by the addition in trans of the T. thermophilus lysS gene despite the fact that this gene was overexpressed and that its product specifically aminoacylates E. coli tRNA(Lys) in vitro.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Chen J,Brevet A,Lapadat-Tapolsky M,Blanquet S,Plateau Pdoi
10.1128/jb.176.9.2699-2705.1994subject
Has Abstractpub_date
1994-05-01 00:00:00pages
2699-705issue
9eissn
0021-9193issn
1098-5530journal_volume
176pub_type
杂志文章abstract::The levels of reduced and oxidized nicotinamide adenine dinucleotides were determined in Xanthobacter flavus during a transition from heterotrophic to autotrophic growth. Excess reducing equivalents are rapidly dissipated following induction of the Calvin cycle, indicating that the Calvin cycle serves as a sink for ex...
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doi:10.1128/JB.112.1.315-326.1972
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/JB.133.2.780-785.1978
更新日期:1978-02-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.117.2.775-782.1974
更新日期:1974-02-01 00:00:00
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journal_title:Journal of bacteriology
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journal_title:Journal of bacteriology
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pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/jb.172.5.2259-2266.1990
更新日期:1990-05-01 00:00:00
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pub_type: 杂志文章
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更新日期:1995-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1982-05-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:1995-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.112.2.797-805.1972
更新日期:1972-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
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更新日期:2005-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2006-05-01 00:00:00