TGFβ1-(78-109)-HBc融合蛋白的原核表达

Prokaryotic expression of TGFβ1-(78-109)-HBV core fusion protein

目的 应用基因工程技术制备TGFβ1 (78- 10 9) HBc融合蛋白 ,作为TGFβ1疫苗 ,用于抗纤维化研究。方法 合成TGFβ1 (78- 10 9)编码区DNA ,连接于质粒载体 ,再在其下游连接HBc编码区 ,测序证实后 ,进行原核表达 ,表达产物以SDS PAGE、ELISA等方法鉴定。结果 经DNA序列分析证实融合基因序列完全正确 ,SDS PAGE证实原核表达产物相对分子质量为 2 5kD ,ELISA检测证实TGFβ1 (78- 10 9)和HBc的抗原表位均可正确暴露。结论 本研究成功构建了TGFβ1 (78- 10 9) HBc融合基因 ,进行了原核表达 ,并初步证实了表达产物的抗原性 。

Objective To construct TGFβ1 32 (78-109) HBV core fusion protein with genetic engineering technology for the future use of this protein as TGF1 vaccine to inhibit fibrosis. Methods TGFβ1 32 (78-109) coding region was synthesized and cloned into the cloning vector. HBV core coding DNA was then inserted into this vector downstream of TGFβ1 32 (78-109) coding region. After the sequence was confirmed by DNA sequencing, the prokaryotic expression was carried out. The expression product was verified by SDS PAGE and ELISA. Results The sequence of TGFβ1 32 (78-109) HBV core fusion gene was correct. The molecular weight of the expression product was approximately 25?kD in SDS PAGE. The exposure of the antigenic epitopes of TGFβ1 32 (78-109) and HBV core was confirmed by ELISA. Conclusion The successful construction of the prokaryotic expression vector of TGFβ1 32 (78-109) HBV core fusion protein and the verification of the antigenecity of the expression product lay the foundation for further research on the inhibition of fibrosis by TGFβ1 vaccine.