Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1989 Apr;55(4):893–896. doi: 10.1128/aem.55.4.893-896.1989

Cloning and expression in Escherichia coli of a xylanase gene from Bacteroides ruminicola 23.

T R Whitehead 1, R B Hespell 1
PMCID: PMC184220  PMID: 2658806

Abstract

A gene coding for xylanase activity in the ruminal bacterial strain 23, the type strain of Bacteroides ruminicola, was cloned into Escherichia coli JM83 by using plasmid pUC18. AB. ruminicola 23 genomic library was prepared in E. coli by using BamHI-digested DNA, and transformants were screened for xylanase activity on the basis of clearing areas around colonies grown on Remazol brilliant blue R-xylan plates. Six clones were identified as being xylanase positive, and all six contained the same 5.7-kilobase genomic insert. The gene was reduced to a 2.7-kilobase DNA fragment. Xylanase activity produced by the E. coli clone was found to be greater than that produced by the original B. ruminicola strain. Southern hybridization analysis of genomic DNA from the related B. ruminicola strains, D31d and H15a, by using the strain 23 xylanase gene demonstrated one hybridizing band in each DNA.

Full text

PDF
896

Images in this article

Selected References

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

  1. BERNS K. I., THOMAS C. A., Jr ISOLATION OF HIGH MOLECULAR WEIGHT DNA FROM HEMOPHILUS INFLUENZAE. J Mol Biol. 1965 Mar;11:476–490. doi: 10.1016/s0022-2836(65)80004-3. [DOI] [PubMed] [Google Scholar]
  2. Bernier R., Jr, Driguez H., Desrochers M. Molecular cloning of a Bacillus subtilis xylanase gene in Escherichia coli. Gene. 1983 Dec;26(1):59–65. doi: 10.1016/0378-1119(83)90036-7. [DOI] [PubMed] [Google Scholar]
  3. Bladen H. A., Bryant M. P., Doetsch R. N. A Study of Bacterial Species from the Rumen Which Produce Ammonia from Protein Hydrolyzate. Appl Microbiol. 1961 Mar;9(2):175–180. doi: 10.1128/am.9.2.175-180.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Dehority B. A. Characterization of several bovine rumen bacteria isolated with a xylan medium. J Bacteriol. 1966 May;91(5):1724–1729. doi: 10.1128/jb.91.5.1724-1729.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dehority B. A. Pectin-fermenting bacteria isolated from the bovine rumen. J Bacteriol. 1969 Jul;99(1):189–196. doi: 10.1128/jb.99.1.189-196.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hespell R. B., Wolf R., Bothast R. J. Fermentation of xylans by Butyrivibrio fibrisolvens and other ruminal bacteria. Appl Environ Microbiol. 1987 Dec;53(12):2849–2853. doi: 10.1128/aem.53.12.2849-2853.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Panbangred W., Kawaguchi O., Tomita T., Shinmyo A., Okada H. Isolation of two beta-xylosidase genes of Bacillus pumilus and comparison of their gene products. Eur J Biochem. 1984 Jan 16;138(2):267–273. doi: 10.1111/j.1432-1033.1984.tb07911.x. [DOI] [PubMed] [Google Scholar]
  9. Sipat A., Taylor K. A., Lo R. Y., Forsberg C. W., Krell P. J. Molecular cloning of a xylanase gene from Bacteroides succinogenes and its expression in Escherichia coli. Appl Environ Microbiol. 1987 Mar;53(3):477–481. doi: 10.1128/aem.53.3.477-481.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  11. Stahl D. A., Flesher B., Mansfield H. R., Montgomery L. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology. Appl Environ Microbiol. 1988 May;54(5):1079–1084. doi: 10.1128/aem.54.5.1079-1084.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES