Construction of eukaryotic expression plasmids of hepatitis B surface antigen and helper T lymphocyte epitope.
BACKGROUND
DNA immunization provides a promising approach to elicit protective humoral and cellular immune responses against HBV. This study was to construct an eukaryotic expression plasmid containing helper T lymphocyte epitope, which will enhance the immunogenicity of a novel hepatitis B virus (HBV) fusion protein DNA vaccine.


METHODS
The target gene containing pan-DR helper T cell epitopes (PADRE) and HBsAg was amplified by polymerase chain reaction (PCR). The PCR products were linked with PMD-18T vector. Plasmid DNA was purified from transformed E.coli competent cell JM109 and digested with Hind III and EcoR I. Then, the target gene was cloned in pcDNA3.1(+) digested by Hind III and EcoR I. Finally, the identity of DNA was verified by digestion and DNA sequencing.


RESULTS
The recombinant expression vectors of pcDNA3.1(+)-PADRE/HBs were identified by restriction enzyme digestion and DNA sequencing. The insert DNA fragment was consistent with the expected sequence.


CONCLUSIONS
The constructed eukaryotic expression plasmid of pcDNA3.1(+)-PADRE/HBs is convenient for further study of eukaryotic transfection and response for cellular and humoral immunity against HBV.
Piao
Yun-Feng
YF
Department of Gastroenterology, First Hospital of Jilin University, Changchun 130021, China.
Tang
Tong-Yu
TY
Niu
Jun-Qi
JQ
Wang
Feng
F
D018984:Epitopes, T-Lymphocyte / Q000235:genetics*; D005057:Eukaryotic Cells / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology*; D007109:Immunity / Q000502:physiology; D010957:Plasmids / Q000096:biosynthesis*; D006377:T-Lymphocytes, Helper-Inducer / Q000502:physiology; D014162:Transfection / Q000379:methods; D014761:Viral Hepatitis Vaccines / Q000235:genetics / Q000276:immunology*
D018984:Epitopes, T-Lymphocyte; D006514:Hepatitis B Surface Antigens; D014761:Viral Hepatitis Vaccines