Abstract:
:Artificially designed gelatins comprising tandemly repeated 30-amino-acid peptide units derived from human alphaI collagen were successfully produced with a Bacillus brevis system. The DNA encoding the peptide unit was synthesized by taking into consideration the codon usage of the host cells, but no clones having a tandemly repeated gene were obtained through the above-mentioned strategy. Minirepeat genes could be selected in vivo from a mixture of every possible sequence encoding an artificial gelatin by randomly ligating the mixed sequence unit and transforming it into Escherichia coli. Larger repeat genes constructed by connecting minirepeat genes obtained by in vivo selection were also stable in the expression host cells. Gelatins derived from the eight-unit and six-unit repeat genes were extracellularly produced at the level of 0.5 g/liter and easily purified by ammonium sulfate fractionation and anion-exchange chromatography. The purified artificial gelatins had the predicted N-terminal sequences and amino acid compositions and a solgel property similar to that of the native gelatin. These results suggest that the selection of a repeat unit sequence stable in an expression host is a shortcut for the efficient production of repetitive proteins and that it can conveniently be achieved by the in vivo selection method. This study revealed the possible industrial application of artificially designed repetitive proteins.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Kajino T,Takahashi H,Hirai M,Yamada Ydoi
10.1128/aem.66.1.304-309.2000keywords:
subject
Has Abstractpub_date
2000-01-01 00:00:00pages
304-9issue
1eissn
0099-2240issn
1098-5336journal_volume
66pub_type
杂志文章abstract::Bacterioplankton are fundamental components of marine ecosystems and influence the entire biosphere by contributing to the global biogeochemical cycles of key elements. Yet, there is a significant gap in knowledge about their diversity and specific activities, as well as environmental factors that shape their communit...
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pub_type: 杂志文章
doi:10.1128/AEM.34.4.371-376.1977
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更新日期:1985-10-01 00:00:00
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:1989-05-01 00:00:00
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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
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pub_type: 杂志文章
doi:10.1128/AEM.60.6.1974-1977.1994
更新日期:1994-06-01 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:2002-01-01 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:2005-04-01 00:00:00