Abstract:
:The Arctic plant growth-promoting rhizobacterium Pseudomonas putida GR12-2 secretes an antifreeze protein (AFP) that promotes survival at subzero temperatures. The AFP is unusual in that it also exhibits a low level of ice nucleation activity. A DNA fragment with an open reading frame encoding 473 amino acids was cloned by PCR and inverse PCR using primers designed from partial amino acid sequences of the isolated AFP. The predicted gene product, AfpA, had a molecular mass of 47.3 kDa, a pI of 3.51, and no previously known function. Although AfpA is a secreted protein, it lacked an N-terminal signal peptide and was shown by sequence analysis to have two possible secretion systems: a hemolysin-like, calcium-binding secretion domain and a type V autotransporter domain found in gram-negative bacteria. Expression of afpA in Escherichia coli yielded an intracellular 72-kDa protein modified with both sugars and lipids that exhibited lower levels of antifreeze and ice nucleation activities than the native protein. The 164-kDa AFP previously purified from P. putida GR12-2 was a lipoglycoprotein, and the carbohydrate was required for ice nucleation activity. Therefore, the recombinant protein may not have been properly posttranslationally modified. The AfpA sequence was most similar to cell wall-associated proteins and less similar to ice nucleation proteins (INPs). Hydropathy plots revealed that the amino acid sequence of AfpA was more hydrophobic than those of the INPs in the domain that forms the ice template, thus suggesting that AFPs and INPs interact differently with ice. To our knowledge, this is the first gene encoding a protein with both antifreeze and ice nucleation activities to be isolated and characterized.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Muryoi N,Sato M,Kaneko S,Kawahara H,Obata H,Yaish MW,Griffith M,Glick BRdoi
10.1128/JB.186.17.5661-5671.2004keywords:
subject
Has Abstractpub_date
2004-09-01 00:00:00pages
5661-71issue
17eissn
0021-9193issn
1098-5530pii
186/17/5661journal_volume
186pub_type
杂志文章abstract::Two methods of mutagenesis, chemical alkylation and insertion of the tetracycline resistance transposon Tn10, were used to generate mutants of Salmonella typhimurium which had reduced levels of endodeoxyribonucleolytic activity. The chemically induced mutations defined a locus, endA, which was cotransduced with serA a...
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pub_type: 杂志文章
doi:10.1128/JB.144.1.173-178.1980
更新日期:1980-10-01 00:00:00
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更新日期:1991-02-01 00:00:00
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doi:10.1128/jb.170.9.3946-3952.1988
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更新日期:1999-05-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/JB.111.3.801-813.1972
更新日期:1972-09-01 00:00:00
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更新日期:2008-06-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/jb.175.23.7697-7701.1993
更新日期:1993-12-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/jb.169.4.1460-1468.1987
更新日期:1987-04-01 00:00:00
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更新日期:2007-02-01 00:00:00
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doi:10.1128/JB.106.1.238-242.1971
更新日期:1971-04-01 00:00:00
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更新日期:2004-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.119.1.242-249.1974
更新日期:1974-07-01 00:00:00
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pub_type: 杂志文章
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更新日期:1985-04-01 00:00:00
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更新日期:1999-06-01 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:1999-11-01 00:00:00
abstract::Spendlove, Rex S. (California State Department of Public Health, Berkeley), Edwin H. Lennette, Charles O. Knight, and Jean N. Chin. Production in FL cells of infectious and potentially infectious reovirus. J. Bacteriol. 92:1036-1040. 1966.-A comparative study was made of the development in, and release from, FL cells ...
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pub_type: 杂志文章
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更新日期:1966-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.92.4.925-930.1966
更新日期:1966-10-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.11.5027-5033.1988
更新日期:1988-11-01 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:2000-09-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.172.8.4263-4270.1990
更新日期:1990-08-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.102.1.30-35.1970
更新日期:1970-04-01 00:00:00
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journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.188.3.874-881.2006
更新日期:2006-02-01 00:00:00
abstract::Three polynucleotide phosphorylase mutations, isolated in heavily mutagenized Escherichia coli strains Q7, Q13, and Q27, were characterized after their transfer by P1 transduction to nearly isogenic strains which lack ribonuclease I. Each strain has a different altered form of polynucleotide phosphorylase. One enzyme ...
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pub_type: 杂志文章
doi:10.1128/JB.97.3.1437-1443.1969
更新日期:1969-03-01 00:00:00