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目的:用酵母双杂交技术筛选白细胞与乙型肝炎病毒截短型表面抗原中蛋白(MHBst)结合蛋白．方法:PCR扩增MHBst基因,连接入酵母表达载体pGBKT7中构建诱饵质粒,转化酵母细胞AH109并在其内表达．后与转化了人白细胞文库质粒的酵母细胞Y187进行配合,在营养缺陷型培养基(SD／-Trp-Leu-His-Ade)和X-α-半乳糖(X-α-gal)上进行双重筛选阳性菌落增菌后提出质粒转化入大肠杆菌(DH5α),并经氨苄青霉素抗性筛选,提取单克隆菌落质粒,酶切鉴定,序列测定后进行生物信息学分析．结果:应用酵母双杂交技术筛选出阳性克隆8个,经生物信息学分析,排除读码框架不正确者,最后得到7种已知基因,分别为ADP核糖基因子2种,RAB6相互作用蛋白(RAB6IP1)1种,CCR4-NOT转录复合体亚基10(CCR4- NOT-10)1种,脂多糖蛋白结合蛋白(LBP)1种,醛缩酶B(ALDOB)1种,补体成分3(C3)1种,丙酮酸脱氢酶(PDH)1种．结论:成功克隆出MHBst结合蛋白．
Objective: To screen the MHBst binding protein of leukocyte and hepatitis B virus truncated surface antigen by yeast two - hybrid technique. Methods: MHBst gene was amplified by PCR, ligated into yeast expression vector pGBKT7 to construct bait plasmid, and transformed into yeast AH109. The yeast cell Y187 transformed with the human leukocyte library plasmid was double-stranded on the auxotrophy medium (SD / -Trp-Leu-His-Ade) and X-α-galactose The positive colonies were screened and transformed into Escherichia coli (DH5α). After being screened by ampicillin, the colony plasmid was extracted and identified by restriction enzyme digestion. After sequencing, bioinformatics analysis was carried out. Results: Eight positive clones were screened by yeast two-hybrid technique. After bioinformatics analysis, the correct reading frame was excluded. Seven known genes were obtained, which were ADP ribosomal gene 2, RAB6 interacting protein 1 type of CCR4-NOT transcription complex subunit 10 (CCR4-NOT-10), 1 type of lipopolysaccharide protein binding protein (LBP), 1 type of aldolase B (ALDOB) 1 species, pyruvate dehydrogenase (PDH) 1 species. Conclusion: The MHBst binding protein was successfully cloned.