Item – Theses Canada

OCLC number
30071580
Author
Wang, Dongsheng,1958-
Title
Molecular cloning and nucleotide sequence of a Streptococcus mutans gene encoding biotin carboxyl carrier protein.
Degree
M. Sc. -- University of Toronto, 1992
Publisher
Ottawa : National Library of Canada = Bibliothèque nationale du Canada, 1993.
Description
2 microfiches.
Notes
University Microfilms order no. UMI00396740.
Includes bibliographical references.
Abstract
Using a streptavidin/biotin labelling technique, we have unexpectedly cloned a gene encoding biotin carboxyl carrier protein (BCCP) from a genomic library of S. mutans strain UT041. In colony lifts, the clone reacted positively to the streptavidin-containing detection system, but could not be detected in Southern blot analysis. The protein blot analysis demonstrated that the BCCP in the clone had produced a "false-positive" (DNA probe-independent) reaction by binding to the streptavidin. Since the recognition of the presence of the BCCP in S. mutans resulted from the interference caused by its expression on gene probing, the unexpected finding could serve as a warning to researchers to be aware that the binding of biotin-containing cellular protein to streptavidin can result in false positive clones. Secondly, the cloned gene might be useful for developing a fusion vector that would allow purification of a cloned protein by streptavidin/biotin binding.
ISBN
0315785551
9780315785557