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Heterologous gene expression and tra...
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Ma, Biao.
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Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium./
作者:
Ma, Biao.
面頁冊數:
162 p.
附註:
Source: Dissertation Abstracts International, Volume: 64-04, Section: B, page: 1622.
Contained By:
Dissertation Abstracts International64-04B.
標題:
Biology, Molecular. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3087142
Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium.
Ma, Biao.
Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium.
- 162 p.
Source: Dissertation Abstracts International, Volume: 64-04, Section: B, page: 1622.
Thesis (Ph.D.)--OGI School of Science & Engineering, 2003.
Lignin peroxidase (LiP) and manganese peroxidase (MnP) are the major components of the lignin degradation system in the white-rot fungus <italic> Phanerochaete chrysosporium</italic>. Expression of <italic>P. chrysosporium </italic> LiP-encoding genes and MnP-encoding genes (<italic>mnp</italic>) is activated by nutrient nitrogen depletion. Moreover, expression of <italic> mnp</italic> requires the presence of Mn<super>2+</super>. Both nitrogen and Mn<super>2+</super> levels regulate mnp expression at the transcriptional level.Subjects--Topical Terms:
1017719
Biology, Molecular.
Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium.
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Heterologous gene expression and transcriptional regulation of manganese peroxidase gene expression in the basidiomycete Phanerochaete chrysosporium.
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162 p.
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Source: Dissertation Abstracts International, Volume: 64-04, Section: B, page: 1622.
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Advisers: Peter Zuber; Michael H. Gold.
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Thesis (Ph.D.)--OGI School of Science & Engineering, 2003.
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Lignin peroxidase (LiP) and manganese peroxidase (MnP) are the major components of the lignin degradation system in the white-rot fungus <italic> Phanerochaete chrysosporium</italic>. Expression of <italic>P. chrysosporium </italic> LiP-encoding genes and MnP-encoding genes (<italic>mnp</italic>) is activated by nutrient nitrogen depletion. Moreover, expression of <italic> mnp</italic> requires the presence of Mn<super>2+</super>. Both nitrogen and Mn<super>2+</super> levels regulate mnp expression at the transcriptional level.
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The enhanced green fluorescent protein (GFP) gene (<italic>egfp</italic>) was used as a reporter in <italic>P. chrysosporium</italic> for gene expression driven by the glyceraldehyde-pdehydrogenase gene (<italic>gpd</italic>) promoter and the MnP isozyme 1 gene (<italic>mnp1</italic>) promoter. The 1.1-kb <italic> gpd</italic> promoter or the 1.5-kb <italic>mnp1</italic> promoter was joined with the 5<super>′</super> end of the <italic>egfp</italic> coding region, while the 250-bp <italic>mnp1</italic> 3<super>′</super> untranslated region (UTR) was joined with the 3<super>′</super> end of the <italic> egfp</italic> coding region. Efficient expression of <italic> egfp</italic> was only observed when an additional intron-containing sequence was inserted in the 5<super>′</super> end of the <italic>egfp</italic> transcription unit. The GFP fluorescence levels faithfully reported the activity of both the <italic>gpd</italic> and <italic>mnp1</italic> promoters. The 1.5-kb <italic>mnp1</italic> promoter was sufficient for promoting <italic> egfp</italic> expression only under nitrogen-limited and Mn<super>2+</super>-sufficient conditions, similar to that required for endogenous MnP expression. A series of deletion, replacement and translocation mutations were introduced in this promoter, and their effects on <italic>egfp</italic> expression in response to Mn<super>2+</super> status were examined. A 33-bp <italic>cis</italic>-element residing 521 by upstream of the translation start codon in the <italic>mnp1 </italic> promoter was identified that participates in the Mn<super>2+</super>-dependent regulation of <italic>mnp1</italic> promoter activity. Negative control is exerted on this promoter element under Mn<super>2+</super>-deficient conditions. All mutations constructed in the 1.5-kb <italic>mnp1</italic> promoter did not affect nitrogen-dependent gene regulation.
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An efficient transformation system for <italic>P. chrysosporium</italic> was developed using the bacterial Bialaphos resistance gene (<italic>bar</italic>) as a selectable marker. The plasmid containing the <italic>bar</italic> coding region joined with the <italic>gpd</italic> promoter and the <italic>mnp1 </italic> 3<super>′</super> UTR onfers resistance to phosphinothricin (the active component of Bialaphos) when introduced into the <italic>P. chrysosporium </italic> wild-type strain OGC101. An efficient homologous expression system for <italic>P. chrysosporium</italic> MnP isozyme 1 was constructed using <italic> bar</italic> as a selectable marker.
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http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3087142
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