Abstract:
:The genes involved in gluconate catabolism (gntP and gntK) in Corynebacterium glutamicum are scattered in the chromosome, and no regulatory genes are apparently associated with them, in contrast with the organization of the gnt operon in Escherichia coli and Bacillus subtilis. In C. glutamicum, gntP and gntK are essential genes when gluconate is the only carbon and energy source. Both genes contain upstream regulatory regions consisting of a typical promoter and a hypothetical cyclic AMP (cAMP) receptor protein (CRP) binding region but lack the expected consensus operator region for binding of the GntR repressor protein. Expression analysis by Northern blotting showed monocistronic transcripts for both genes. The expression of gntP and gntK is not induced by gluconate, and the gnt genes are subject to catabolite repression by sugars, such as glucose, fructose, and sucrose, as was detected by quantitative reverse transcription-PCR (qRT-PCR). Specific analysis of the DNA promoter sequences (PgntK and PgntP) was performed using bifunctional promoter probe vectors containing mel (involved in melanin production) or egfp2 (encoding a green fluorescent protein derivative) as the reporter gene. Using this approach, we obtained results parallel to those from qRT-PCR. An applied example of in vivo gene expression modulation of the divIVA gene in C. glutamicum is shown, corroborating the possible use of the gnt promoters to control gene expression. glxR (which encodes GlxR, the hypothetical CRP protein) was subcloned from the C. glutamicum chromosomal DNA and overexpressed in corynebacteria; we found that the level of gnt expression was slightly decreased compared to that of the control strains. The purified GlxR protein was used in gel shift mobility assays, and a specific interaction of GlxR with sequences present on PgntP and PgntK fragments was detected only in the presence of cAMP.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Letek M,Valbuena N,Ramos A,Ordóñez E,Gil JA,Mateos LMdoi
10.1128/JB.188.2.409-423.2006keywords:
subject
Has Abstractpub_date
2006-01-01 00:00:00pages
409-23issue
2eissn
0021-9193issn
1098-5530pii
188/2/409journal_volume
188pub_type
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journal_title:Journal of bacteriology
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更新日期:1988-08-01 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:2010-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JB.142.1.254-261.1980
更新日期:1980-04-01 00:00:00
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pub_type: 杂志文章
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