Abstract:
:A method for the measurement of specific levels of gene expression that combines target mRNA and target DNA quantitation has been developed. The use of target gene dose as a normalizing factor for mRNA provides an alternative to 16S or 23S rRNA, which are unsuitable for use in the environment because of their presence in nontarget organisms. Both target mRNA and DNA are recovered from replicate samples and detected by using antisense and sense single-stranded RNA gene probes. For efficient mRNA recovery, the use of Millipore Durapore filters and multiple extractions was necessary. Quantitation was performed by radiometric detection by using a beta-scanner and comparison of the sample signal against target mRNA and DNA standard curves. This method enabled the measurement of expression of the catechol-2,3-dioxygenase gene (xylE) contained on the thermoregulated plasmid pLV1013 in a marine Vibrio strain in culture and in the environment. In studies of the relationship between mRNA levels and enzyme activities, the appearance of enzyme activity lagged behind xylE mRNA synthesis by an hour after temperature induction. This suggests that mRNA analysis is well suited for determining rapid regulation of microbial gene expression at the transcriptional level in water column microbial populations.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Pichard SL,Paul JHdoi
10.1128/AEM.59.2.451-457.1993keywords:
subject
Has Abstractpub_date
1993-02-01 00:00:00pages
451-7issue
2eissn
0099-2240issn
1098-5336journal_volume
59pub_type
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.67.8.3702-3706.2001
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.45.3.872-877.1983
更新日期:1983-03-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.60.3.932-939.1994
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pub_type: 杂志文章
doi:10.1128/AEM.57.3.642-644.1991
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.50.4.919-923.1985
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journal_title:Applied and environmental microbiology
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pub_type: 杂志文章
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更新日期:2020-12-18 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.62.11.4009-4013.1996
更新日期:1996-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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pub_type: 杂志文章
doi:10.1128/AEM.65.2.540-548.1999
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2018-07-02 00:00:00
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pub_type: 杂志文章
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更新日期:2020-03-02 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:1996-04-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2000-08-01 00:00:00
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pub_type: 杂志文章
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更新日期:2009-06-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.65.11.5117-5123.1999
更新日期:1999-11-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.71.12.8836-8845.2005
更新日期:2005-12-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2009-02-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/aem.66.6.2349-2357.2000
更新日期:2000-06-01 00:00:00