Abstract:
:The Escherichia coli K-12 hisT gene has been cloned, and its organization and expression have been analyzed on multicopy plasmids. The hisT gene, which encodes tRNA pseudouridine synthase I (PSUI), was isolated on a Clarke-Carbon plasmid known to contain the purF gene. The presence of the hisT gene on this plasmid was suggested by its ability to restore both production of PSUI enzymatic activity and suppression of amber mutations in a hisT mutant strain. A 2.3-kilobase HindIII-ClaI restriction fragment containing the hisT gene was subcloned into plasmid pBR322, and the resulting plasmid (designated psi 300) was mapped with restriction enzymes. Complementation analysis with different kinds of hisT mutations and tRNA structural analysis confirmed that plasmid psi 300 contained the hisT structural gene. Enzyme assays showed that plasmid psi 300 overproduced PSUI activity by ca. 20-fold compared with the wild-type level. Subclones containing restriction fragments from plasmid psi 300 inserted downstream from the lac promoter established that the hisT gene is oriented from the HindIII site toward the ClaI site. Other subclones and derivatives of plasmid psi 300 containing insertion or deletion mutations were constructed and assayed for production of PSUI activity and production of proteins in minicells. These experiments showed that: (i) the proximal 1.3-kilobase HindIII-BssHII restriction fragment contains a promoter for the hisT gene and encodes a 45,000-dalton polypeptide that is not PSUI; (ii) the distal 1.0-kilobase BssHII-ClaI restriction fragment encodes the 31,000-dalton PSUI polypeptide; (iii) the 45,000-dalton polypeptide is synthesized in an approximately eightfold excess compared with PSUI; and (iv) synthesis of the two polypeptides is coupled, suggesting that the two genes are part of an operon. Insertion of mini-Mu d1 (lac Km) phage into plasmid psi 300 confirmed that the hisT gene is the downstream gene in the operon.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Marvel CC,Arps PJ,Rubin BC,Kammen HO,Penhoet EE,Winkler MEdoi
10.1128/JB.161.1.60-71.1985subject
Has Abstractpub_date
1985-01-01 00:00:00pages
60-71issue
1eissn
0021-9193issn
1098-5530journal_volume
161pub_type
杂志文章abstract::The Bacillus subtilis SpoVE integral membrane protein is essential for the heat resistance of spores, probably because of its involvement in spore peptidoglycan synthesis. We found that an SpoVE-yellow fluorescent protein (YFP) fusion protein becomes localized to the forespore during the earliest stages of engulfment,...
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doi:10.1128/JB.127.1.595-609.1976
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pub_type: 杂志文章
doi:10.1128/jb.176.9.2759-2762.1994
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.22.5921-5927.1998
更新日期:1998-11-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/jb.178.11.3127-3132.1996
更新日期:1996-06-01 00:00:00
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pub_type: 杂志文章
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更新日期:2007-03-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.164.1.270-275.1985
更新日期:1985-10-01 00:00:00
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journal_title:Journal of bacteriology
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doi:10.1128/JB.183.21.6355-6364.2001
更新日期:2001-11-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.143.2.1090-1094.1980
更新日期:1980-08-01 00:00:00
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更新日期:1999-09-01 00:00:00
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更新日期:1975-06-01 00:00:00
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更新日期:1994-04-01 00:00:00
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更新日期:2015-02-15 00:00:00
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更新日期:2007-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.172.2.949-955.1990
更新日期:1990-02-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1970-03-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1976-04-01 00:00:00
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journal_title:Journal of bacteriology
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更新日期:1999-04-01 00:00:00
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更新日期:2017-02-28 00:00:00
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更新日期:1988-06-01 00:00:00