Document Detail

Characterization, identification, and cloning of the S-layer protein from Cytophaga sp.
MedLine Citation:
PMID:  18322733     Owner:  NLM     Status:  MEDLINE    
We characterized, identified, and cloned a major protein which comprised 16% of the total proteins from Cytophaga sp. cell lysate. After French pressing, the fraction of cell envelope was treated with 0.2% Triton X-100 to remove cell membranes. Subsequent SDS-PAGE analysis of the Triton X-100-insoluble cell wall revealed a protein of 120 kDa with a pI of 5.4, which was identified by gold immunostaining as the surface (S)-layer protein of this soil bacterium. The nucleotide sequence of the cloned S-layer protein gene (slp) encoding this protein consisted of 3144 nucleotides with an ORF for 1047 amino acids, which included a typical 32-amino acid leader peptide sequence. Amino acid sequence alignment revealed 29-48% similarity between this protein and the S-layer proteins from other prokaryotic organisms. The 120-kDa protein from the Cytophaga sp. cell lysate has been characterized as a member of the S-layer proteins, and the slp gene was cloned and expressed in Escherichia coli. E. coli harboring the plasmid containing the 600- or 800-bp DNA fragment upstream of the initiation codon of the slp gene, in the presence of the reporter gene rsda (raw starch digesting amylase), showed amylase activity in starch containing plate. The putative promoter region of slp located 600 bp upstream of the initiation codon might be used for foreign gene expression.
Shiow Ying Chiou; Pei Ling Kang; Tai Way Liao; Chii Ling Jeang
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Publication Detail:
Type:  Journal Article     Date:  2008-03-06
Journal Detail:
Title:  Current microbiology     Volume:  56     ISSN:  0343-8651     ISO Abbreviation:  Curr. Microbiol.     Publication Date:  2008 Jun 
Date Detail:
Created Date:  2008-05-12     Completed Date:  2008-08-14     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  7808448     Medline TA:  Curr Microbiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  597-602     Citation Subset:  IM    
Department of Food Science and Biotechnology, National Chung-Hsing University, 250 Kuo Kuang Road, Taichung, 402, Taiwan, R.O.C.
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MeSH Terms
Cloning, Molecular*
Cytophaga / chemistry,  genetics*,  metabolism,  ultrastructure
Electrophoresis, Polyacrylamide Gel
Gene Expression
Membrane Glycoproteins / chemistry*,  genetics,  isolation & purification,  metabolism
Molecular Sequence Data
Molecular Weight
Open Reading Frames
Promoter Regions, Genetic
Sequence Analysis, DNA
Reg. No./Substance:
0/Membrane Glycoproteins; 0/S-layer proteins

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