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  Molecular cloning, expression, and characterization of the gene for the surface (HPI)-layer protein of Deinococcus radiodurans in Escherichia coli

Peters, J., & Baumeister, W. (1986). Molecular cloning, expression, and characterization of the gene for the surface (HPI)-layer protein of Deinococcus radiodurans in Escherichia coli. Journal of Bacteriology., 167(3), 1048-1054.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-74EC-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-74ED-9
Genre: Journal Article

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Peters, J.1, Author              
Baumeister, W.1, Author              
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1External Organizations, ou_persistent22              

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Free keywords: *Bacterial Proteins/ge [Genetics]; Cloning, Molecular; Escherichia coli/me [Metabolism]; *Genes, Bacterial; Genes, Structural; *Micrococcus/ge [Genetics]; Promoter Regions (Genetics); Recombinant Proteins/bi [Biosynthesis]; Recombinant Proteins/ge [Genetics]; Surface Properties
 Abstract: The HPI protein of Deinococcus radiodurans belongs to the class of surface layer proteins which form crystalline two-dimensional arrays on bacterial cell envelopes. We have cloned and expressed the gene for this protein of Mr about 100,000 by using plasmid pUC8 in Escherichia coli. As judged by immunoreaction with monospecific antibodies, apparent Mr, and limited proteolysis, a single clone contained the gene encoding the complete polypeptide on an 8.9-kilobase (kb) insert. The insert was reduced to a 5.7-kb HindIII fragment, cloned in pUC18 in both orientations, and subjected to unilateral processive deletion with exonuclease III. The library of deletion derivatives was mapped and, in conjunction with protein immunoblotting of expressed polypeptides, was used to locate the positions of the structural gene and the Deinococcus promoter region that was responsible for expression of the HPI polypeptide. The HPI gene was confined to a stretch 2.95 kb in length.

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 Dates: 1986
 Publication Status: Published in print
 Pages: -
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 Rev. Type: -
 Identifiers: eDoc: 318752
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Title: Journal of Bacteriology.
Source Genre: Journal
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Pages: - Volume / Issue: 167 (3) Sequence Number: - Start / End Page: 1048 - 1054 Identifier: -