E3泛素连接酶HECTD3真核表达质粒的构建

摘要

目的：克隆E3泛素连接酶(homologous to the E6-associated protein carboxyl terminus domain containing 3, HECTD3)基因及构建真核表达质粒。方法提取人卵巢癌细胞SKOV-3细胞RNA,并逆转录为cDNA,以此作为模板PCR法扩增HECTD3基因,将其克隆至真核表达载体pcDNA3.1(+),测序验证构建质粒中包含HECTD3基因。结论 HECTD3基因克隆及真核表达质粒构建成功,可以用于后续卵巢癌发生发展的分子生物学研究。%Objective To clone the gene of homologous to the E6-associated protein carboxyl terminus domain containing 3 (HECTD3) and construct the HECTD3 gene expression plasmid. Methods The HECTD3 gene was cloned from ovarian cell cDNA by PCR and recombined into the eukaryotic expression plasmid pcDNA3.1(+). Results the HECTD3 gene was confirmed in the eukaryotic expression plasmid by sequence analysis. Conclusion The HECTD3 gene expression plasmid can be the tools for further research of ovarian cancer.