Glypican3 N端融合基因构建及鉴定  

Construction and Identification of Glypican3 N-terminal Fusion Gene

作　　者：周玉宏[1] 倪润洲[1] 肖明兵[1] 刘海鸥[2] 

机构地区：[1]南通大学附属医院消化病研究室,江苏南通226001 [2]南通大学医学院微生物与免疫学教研室,江苏南通226001

出　　处：《现代检验医学杂志》2008年第1期46-48,共3页Journal of Modern Laboratory Medicine

摘　　要：目的构建人磷酯酰肌醇蛋白聚糖3(glypican3,GPC3)N端融合基因。方法采用RT-PCR技术,从人肝癌细胞株HepG2 mRNA中扩增GPC3 N端基因片段,经双酶切后,T4DNA连接酶将此片段定向连接至质粒pcDNA 3.1,并转化大肠埃希菌DH5α,增菌培养后提取质粒并PCR扩增、酶切鉴定及序列测定。结果GPC3 N端基因片段正确插入质粒pcDNA 3.1,片段大小及序列正确。结论成功构建重组质粒pcDNA 3.1-GPC3 N。Objective To construct human glypican3 N-terminal fusion gene. Methods The glypican3 N-terminal gene was amplified by RT-PCR from human HCC HepG2 cell line mRNA. The RT-PCR product was ligated into plasmid pcDNA3. 1 by double digestion and T4 DNA ligase. The recombinant plasmid was transformed into E. coli DH5α and then amplified by PCR. Results Glypican3 N-terminal gene was correctly inserted into plasmid pcDNA3.1,which was confirmed by both identification with enzyme digestion and DNA sequencing. The DNA sequence was 100% homogeneous with that of glypican3 reported. Conclusion Recombinant plasmid pcDNA3.1-glypican3 N was constructed.

关 键 词：glypican3 质粒 融合基因构建 

分 类 号：Q78[生物学—分子生物学]