Cloning, sequencing, and expression in Escherichia coli of a Streptococcus faecalis autolysin.
A Streptococcus faecalis genomic bank was obtained by partial digestion with MboI and cloning into the SalI restriction site of pTZ18R. Screening of about 60,000 Escherichia coli transformants for cell wall lysis activity was done by exposing recombinant colonies grown on medium containing lyophilized Micrococcus lysodeikticus cells to chloroform and toluene vapors in order to release proteins. Because this procedure provoked cell death, colonies could not be used directly for transformant recovery; however, recovery was achieved by partial purification of plasmid DNA from active colonies on the agar plate and transformation of E. coli competent cells. About 60 recombinants were found. One of them (pSH6500) codes for a lytic enzyme active against S. faecalis and M. lysodeikticus cell walls. A shorter clone (pSH4000) was obtained by deleting an EcoRI fragment from the 6.5-kb original insert, leaving a 4-kb EcoRI-MboI insert; this subclone expressed the same lytic activity. Sequencing of a portion of pSH4000 revealed a unique open reading frame of 2,013 nucleotides coding for a 641-amino-acid (74-kDa) polypeptide and containing four 204-nucleotide direct repeats.
27 references, click to expand
-
3'-end modifications of the Streptococcus pneumoniae lytA gene: role of the carboxy terminus of the pneumococcal autolysin in the process of enzymatic activation (conversion).Gene. 1987;61(1):13-9 PMID: 2895040
-
The second peptidoglycan hydrolase of Streptococcus faecium ATCC 9790 covalently binds penicillin.J Bacteriol. 1989 Aug;171(8):4355-61 PMID: 2753858
-
Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements.J Mol Biol. 1983 Jun 5;166(4):477-535 PMID: 6864790
-
The penicillin-binding proteins in Streptococcus faecalis ATCC 9790.Eur J Biochem. 1980 Sep;110(2):445-56 PMID: 6777159
-
Covalent modification of the beta-1,4-N-acetylmuramoylhydrolase of Streptococcus faecium with 5-mercaptouridine monophosphate.Proc Natl Acad Sci U S A. 1988 Sep;85(18):6667-71 PMID: 3413115
-
Nucleotide sequence of the lysozyme gene of bacteriophage T4. Analysis of mutations involving repeated sequences.J Mol Biol. 1983 Apr 5;165(2):229-48 PMID: 6302287
-
Identification of a streptococcal penicillin-binding protein that reacts very slowly with penicillin.J Bacteriol. 1983 Sep;155(3):1343-50 PMID: 6411688
-
Studies on transformation of Escherichia coli with plasmids.J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
-
A rapid alkaline extraction method for the isolation of plasmid DNA.Methods Enzymol. 1983;100:243-55 PMID: 6353143
-
A comprehensive set of sequence analysis programs for the VAX.Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95 PMID: 6546423
-
DNA sequencing with chain-terminating inhibitors.Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
-
Use of bacteriolytic enzymes in determination of wall structure and their role in cell metabolism.Bacteriol Rev. 1968 Dec;32(4 Pt 2):425-64 PMID: 4884715
-
The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342-6 PMID: 4598299
-
"In vivo" detection of L-arabinose-binding protein, CRM-negative mutants.J Bacteriol. 1971 Feb;105(2):604-8 PMID: 5541535
-
Molecular evolution of lytic enzymes of Streptococcus pneumoniae and its bacteriophages.Proc Natl Acad Sci U S A. 1988 Feb;85(3):914-8 PMID: 3422470
-
Nucleotide sequence of the small double-stranded RNA segment of bacteriophage phi 6: novel mechanism of natural translational control.J Virol. 1986 Apr;58(1):142-51 PMID: 3754015
-
Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli.Gene. 1986;43(3):265-72 PMID: 2875013
-
'DNA Strider': a 'C' program for the fast analysis of DNA and protein sequences on the Apple Macintosh family of computers.Nucleic Acids Res. 1988 Mar 11;16(5):1829-36 PMID: 2832831
-
An improved technique for the efficient construction of gene libraries by partial filling-in of cohesive ends.Gene. 1986;42(1):119-23 PMID: 3013727
-
Nucleotide sequence of Bacillus phage phi 29 genes 14 and 15: homology of gene 15 with other phage lysozymes.Nucleic Acids Res. 1986 Dec 22;14(24):10001-8 PMID: 3027653
-
Phage P22 lysis genes: nucleotide sequences and functional relationships with T4 and lambda genes.Virology. 1985 May;143(1):280-9 PMID: 2998005
-
Nucleotide sequence of the late region of Bacillus subtilis phage PZA, a close relative of phi 29.Gene. 1986;44(1):107-14 PMID: 3095188
-
Cloning, sequencing and expression of a Bacillus bacteriolytic enzyme in Escherichia coli.Mol Gen Genet. 1988 Oct;214(2):241-8 PMID: 3070348
-
Turnover of cell walls in microorganisms.Microbiol Rev. 1988 Dec;52(4):554-67 PMID: 3070324
-
Purification and characterization of the autolytic glycosidase of Streptococcus pneumoniae.Biochem Biophys Res Commun. 1989 Jan 16;158(1):251-6 PMID: 2492193
-
Detection of bacterial cell wall hydrolases after denaturing polyacrylamide gel electrophoresis.Can J Microbiol. 1989 Aug;35(8):749-53 PMID: 2819603
-
Paradoxical response of Enterococcus faecalis to the bactericidal activity of penicillin is associated with reduced activity of one autolysin.Antimicrob Agents Chemother. 1990 Feb;34(2):314-20 PMID: 2109578
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