β-淀粉样肽与乙型肝炎核心抗原融合基因Aβ-HBcAg表达载体的构建

目的　构建 β 淀粉样肽 ( β amyloidpeptide ,Aβ)与乙肝核心抗原 (HBcAg)融合基因Aβ HBcAg的原核表达载体 pBV2 2 0 /Aβ HBcAg ,为阿尔茨海默病基因工程疫苗的研究和制备奠定基础。 方法　采用非对称互补引物 /模板法 ,制备两端含有酶切位点的Aβ1- 42肽的cDNA ,将其连接到删除了Pre C区的HBcAg亚型ayw基因的N端组成融合基因Aβ HBcAg ,再将该融合基因亚克隆于载体 pBV2 2 0 ,构建为原核表达载体pBV2 2 0 /Aβ HBcAg。 结果　经DNA测序 ,限制性内切酶酶切等方法证实已成功地将Aβ1- 42cDNA重组到HBVcore的N端 ,并将融合基因亚克隆于 pBV2 2 0内。 结论　成功构建了原核表达载体 pBV2 2 0 /Aβ HBcAg。

Construction of expression vector for Aβ-HBcAg fusion gene

Objective To construct prokaryotic expression vector bearing Aβ-HBcAg fusion gene and lay foundation for the study and manufacturing of genetic engineering vaccine of Alzheimer's disease.Methods By means of asymmetrical complement primer/template, double stranded cDNA of Aβ1-42 was gained. The fusion gene named Aβ-HBcAg was obtained by ligating Aβ1-42 cDNA and ayw. one subtype of HBcAg, which pre-C domain was deleted. Then it was subcloned into prokaryotic expression vector pBV220. Results Evidences of DNA sequence analysis and restriction enzymes digestion showed that we recombined Aβ1-42 cDNA to the N-terminal of ayw gene, and the fusion gene was subcloned into pBV220 successfully.Conclusion Prokaryotic expression vector pBV220/Aβ-HBcAg can be constructed successfully.