Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jun;173(12):3918–3920. doi: 10.1128/jb.173.12.3918-3920.1991

Overproduction and purification of McrC protein from Escherichia coli K-12.

L Zheng 1, H D Braymer 1
PMCID: PMC208028  PMID: 2050643

Abstract

The McrC protein, encoded by one of the two genes involved in the McrB restriction system, was produced in Escherichia coli cells by using a T7 expression system. Following sequential DEAE-Sepharose and hydroxylapatite column chromatography, the protein was purified to apparent homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of the purified McrC protein agreed exactly with the one deduced from the DNA sequence by Ross et al. (J. Bacteriol. 171:1974-1981, 1989).

Full text

PDF
3918

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Butkus V., Klimasauskas S., Petrauskiene L., Maneliene Z., Lebionka A., Janulaitis A. Interaction of AluI, Cfr6I and PvuII restriction-modification enzymes with substrates containing either N4-methylcytosine or 5-methylcytosine. Biochim Biophys Acta. 1987 Aug 25;909(3):201–207. doi: 10.1016/0167-4781(87)90078-9. [DOI] [PubMed] [Google Scholar]
  2. Dila D., Sutherland E., Moran L., Slatko B., Raleigh E. A. Genetic and sequence organization of the mcrBC locus of Escherichia coli K-12. J Bacteriol. 1990 Sep;172(9):4888–4900. doi: 10.1128/jb.172.9.4888-4900.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Raleigh E. A., Trimarchi R., Revel H. Genetic and physical mapping of the mcrA (rglA) and mcrB (rglB) loci of Escherichia coli K-12. Genetics. 1989 Jun;122(2):279–296. doi: 10.1093/genetics/122.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Raleigh E. A., Wilson G. Escherichia coli K-12 restricts DNA containing 5-methylcytosine. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9070–9074. doi: 10.1073/pnas.83.23.9070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ross T. K., Achberger E. C., Braymer H. D. Characterization of the Escherichia coli modified cytosine restriction (mcrB) gene. Gene. 1987;61(3):277–289. doi: 10.1016/0378-1119(87)90191-0. [DOI] [PubMed] [Google Scholar]
  6. Ross T. K., Achberger E. C., Braymer H. D. Identification of a second polypeptide required for McrB restriction of 5-methylcytosine-containing DNA in Escherichia coli K12. Mol Gen Genet. 1989 Apr;216(2-3):402–407. doi: 10.1007/BF00334382. [DOI] [PubMed] [Google Scholar]
  7. Ross T. K., Achberger E. C., Braymer H. D. Nucleotide sequence of the McrB region of Escherichia coli K-12 and evidence for two independent translational initiation sites at the mcrB locus. J Bacteriol. 1989 Apr;171(4):1974–1981. doi: 10.1128/jb.171.4.1974-1981.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ross T. K., Braymer H. D. Localization of a genetic region involved in McrB restriction by Escherichia coli K-12. J Bacteriol. 1987 Apr;169(4):1757–1759. doi: 10.1128/jb.169.4.1757-1759.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Sain B., Murray N. E. The hsd (host specificity) genes of E. coli K 12. Mol Gen Genet. 1980;180(1):35–46. doi: 10.1007/BF00267350. [DOI] [PubMed] [Google Scholar]
  10. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  11. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES