[Cloning and sequence analysis of recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit and Actinobacillus actinomycetemcomitans fimbria associative protein].
OBJECTIVE
To clone the recombinant fusion gene of Escherichia coli heat-liable enterotoxin B subunit (Ltb) and Actinobacillus actinomycetemcomitans fimbria associative protein (Fap).


METHODS
Two 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.


RESULTS
The ltb-fap about 531bp in size was obtained successfully, and identified by PCR, restrictive enzyme and sequence analysis.


CONCLUSIONS
The vector of pET28a ltb-fap was obtained.
Li
Yi
Y
Dept. of Oral Medicine, School of Stomatology, Jilin University, Changchun, China.
Sun
Hong-chen
HC
Guo
Xue-jun
XJ
Feng
Shu-zhang
SZ
D016976:Aggregatibacter actinomycetemcomitans; D001427:Bacterial Toxins; D003001:Cloning, Molecular; D019976:Cloning, Organism; D004768:Enterotoxins; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D006358:Hot Temperature; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins; D017421:Sequence Analysis
D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins