一种新型HBsAg真核表达载体的构建

目的　在CHO细胞中高效表达HBsAg。方法　通过PCR方法扩增出S和dhfr基因片段 ,分别克隆到T载体上。然后分别将S和dhfr基因克隆到pCI载体上 ,构建pCI S dhfr真核表达载体。转染CHO(dhfr )细胞 ,并用ELISA方法检测HBsAg表达量。结果　S和dhfr基因经测序与Genbank报道一致。PCR和测序结果与预期一致。转染后检测结果呈阳性。结论　已成功构建在CHO(dhfr )细胞中高效表达的pCI S dhfr双顺反子真核表达载体

Construction of A Novel Eukaryotic Expression Vector of HBsAg

Objective To highly express HBsAg in CHO cells.Methods Amplify S and dhfr gene fragments by PCR and insert into T vector respectively, then sub-clone into pCI vector to construct an eukaryotic expression vector pCI-S-dhfr. Transfect CHO(dhfr-) cells with the vector pCI-S-dhfr and determine the expression level of HBsAg with its corresponding diagnostic kit.Results The sequences of S and dhfr genes were consistent with those reported in GenBank. The results of PCR and sequencing of fusion gene pCI-S-dhfr were also consistent with those expected. The expressed HBsAg was detected in the transfected cells.Conclusion A dicistronic expression vector pCI-S-dhfr was successfully constructed, and HBsAg was highly expressed in CHO(dhfr-)cells.