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Titolo:
CLONING AND SEQUENCE-ANALYSIS OF A HEMOLYSIN-ENCODING GENE FROM PSEUDOMONAS-PAUCIMOBILIS
Autore:
MINNICK MF; SCHERER DC;
Indirizzi:
UNIV MONTANA,DIV BIOL SCI MISSOULA MT 59812
Titolo Testata:
Gene
fascicolo: 1, volume: 130, anno: 1993,
pagine: 57 - 63
SICI:
0378-1119(1993)130:1<57:CASOAH>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
ESCHERICHIA-COLI; NUCLEOTIDE-SEQUENCE; MOLECULAR-CLONING; ALPHA-HEMOLYSIN; INFECTIONS; AERUGINOSA; VIRULENCE; CLEAVAGE; PROTEINS;
Keywords:
CYTOLYSIN; RECOMBINANT DNA; NUCLEOTIDE SEQUENCE; PSEUDOMONAD; OPPORTUNISTIC PATHOGEN;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
44
Recensione:
Indirizzi per estratti:
Citazione:
M.F. Minnick e D.C. Scherer, "CLONING AND SEQUENCE-ANALYSIS OF A HEMOLYSIN-ENCODING GENE FROM PSEUDOMONAS-PAUCIMOBILIS", Gene, 130(1), 1993, pp. 57-63

Abstract

We report the cloning, expression and nucleotide (nt) sequence of a beta-hemolysin-encoding gene, termed hlyA, from Pseudomonas paucimobilis. A genomic DNA library of the pseudomonad was constructed in Escherichia coli using the plasmid vector, pUC19. The hlyA gene was cloned byscreening for a beta-hemolytic phenotype in E. coli transformants andwas mapped to a 1100-bp PstI-SmaI fragment. The nt sequence analysis of the 1100-bp insert revealed a 789-bp open reading frame which is preceded by a 10-nt purine-rich sequence with a possible ribosome-binding site of GGA. The ORF terminates with a single UGA stop codon and is immediately followed by a large inverted repeat with 27-bp arms which may serve as a Rho-factor-independent transcriptional terminator. The hlyA gene codes for a protein of 263 amino acids (aa) residues with a deduced relative molecular mass (M(r)) of 29 695 and a predicted pI value of 11.5. Expression of hlyA from recombinant DNA in E. coli occurred regardless of insert orientation in the vector and produced a 29-kDa protein. Confirmation of P. paucimobilis as the source of the clonedhlyA gene was determined by DNA hybridization. A search of various ntand aa sequence databases revealed no homologues to hlyA or its encoded protein.

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Documento generato il 08/07/20 alle ore 08:13:25