Cloning and sequence analysis of recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit and Actinobacillus actinomycetemcomitans fimbria associative protein.
- Author:
Yi LI
1
;
Hong-chen SUN
;
Xue-jun GUO
;
Shu-zhang FENG
Author Information
- Publication Type:Journal Article
- MeSH: Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Cloning, Molecular; Cloning, Organism; Enterotoxins; Escherichia coli; Escherichia coli Proteins; Hot Temperature; Plasmids; Polymerase Chain Reaction; Recombinant Fusion Proteins; Sequence Analysis
- From: West China Journal of Stomatology 2005;23(1):24-40
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone the recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit (Ltb) and Actinobacillus actinomycetemcomitans fimbria associative protein (Fap).
METHODSTwo couples of primers were designed for PCR according to the known sequence of ltb and fap. The ltb and fap gene were obtained by amplification PCR technique from plasmid EWD299 of Escherichia coli and Actinobacillus actinomycetemcomitans 310 DNA respectively, and fused them by PCR. The fusion gene ltb-fap were cloning into plasmid pET28a(+). The recombined plasmid pET28a ltb-fap was transformed into Escherichia coli DH5alpha. The recombinant was screened and identified by restriction enzyme and PCR. The cloned gene was sequenced.
RESULTSThe ltb-fap about 531bp in size was obtained successfully, and identified by PCR, restrictive enzyme and sequence analysis.
CONCLUSIONThe vector of pET28a ltb-fap was obtained.
