[Cloning and sequencing of fusion protein gene of recombinant GM-HBsAg].
Junli Xue, Qi Chen, Qing Luo
Abstract
Junli Xue, Qi Chen, Qing Luo
Abstract
GM-CSF is an effective adjuvant for immunology, it was used to improve the immunological function of antigen and to increase the anti-HBs responses. Through using polymerase chain reaction (PCR) technique, we amplified 384 bp and 846 bp gene fragments from GM-CSF and HBsAg. The products were extracted and purified; a 1230 bp gene fragment was obtained by T4 DNA linkage. This amplified product was cloned into pUC19 vector. The positive clones were screened and the inserted fragment was determined by restriction enzymes. The gene fragment length determined is 1230 bp, the sequence analysis showed that the gene fragment is GM-CSF-HBsAg gene. The successful construction of GM-HBsAg fusion protein gene provides basis for research of its biological activity and expression.
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GM-CSF is an effective adjuvant for immunology, it was used to improve the immunological function of antigen and to increase the anti-HBs responses. Through using polymerase chain reaction (PCR) technique, we amplified 384 bp and 846 bp gene fragments from GM-CSF and HBsAg. The products were extracted and purified; a 1230 bp gene fragment was obtained by T4 DNA linkage. This amplified product was cloned into pUC19 vector. The positive clones were screened and the inserted fragment was determined by restriction enzymes. The gene fragment length determined is 1230 bp, the sequence analysis showed that the gene fragment is GM-CSF-HBsAg gene. The successful construction of GM-HBsAg fusion protein gene provides basis for research of its biological activity and expression.
Key concepts: Molecular biology, Restriction enzyme, Gene, Recombinant DNA, Fusion gene, Fusion protein, pUC19, Biology