[Construction of a dicistronic expression plasmid vector containing double-valent hepatitis B surface gene].
OBJECTIVE
In order to enhance vaccine response, we constructed a dicistronic expression plasmid containing double HBsAg immunogenes.


METHODS
At first, pcDNA3.1 plasmid vector was digested with NheI and EcoRI to get the coding sequence of the small (S) surface protein of HBV, then cloned into pCI-neo vector and named it pCI-S. By PCR amplification, the product of IRES-S was digested with SalI & BamHI, and cloned into pBluescript IIK+S to generate pBKS-IRES-S vector, then subcloned to the pCI-S plasmid to generate pCI-S-IRES-S, which is a dicistronic plasmid of double value HBsAg genes.


RESULTS
Two plasmids we constructed were digested with related restriction nucleic enzymes. Sequence analysis of HBsAg and IRES-S gene did not reveal any mutation.


CONCLUSIONS
The construction of dicistronic plasmid of divalue HBsAg immunogenes has been well cloned, which is convenient for further research on cell expression and gene immunization in animals.
Wang
B
B
Institute for Vrial Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Zhang
D
D
Huang
A
A
Yao
Y
Y
Liu
Q
Q
Ren
H
H
D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping
D000956:Antigens, Viral; D006514:Hepatitis B Surface Antigens