E3泛素连接酶HECTD3真核表达质粒的鉴定

摘要

目的：鉴定E3泛素连接酶(homologous to the E6-associated protein carboxyl terminus domain containing 3，HECTD3)基因的真核表达质粒。方法提取无内毒素真核表达质粒pcDNA-HECTD3，利用lipofectamine 2000将其转染进卵巢癌细胞SKOV-3细胞，qRT-PCR法和蛋白质免疫印迹技术检测该基因的表达情况。结果真核表达质粒pcDNA-HECTD3可以显著地提高HECTD3基因在mRNA和蛋白水平的表达量。结论 HECTD3基因的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 confirm the eukaryotic expression plasmid containing the gene of homologous to the E6-associated protein carboxyl terminus domain containing 3 (HECTD3). Methods Extracted the endo-free plasmid pcDNA-HECTD3 and transfected it into ovarian cellSKOV-3, final y qRT-PCR and western blot assay were used to measure HECTD3 expression levels. Results HECTD3 gene was remarkably overexpressed in transfected cells. Conclusion The HECTD3 gene expression plasmid can be the tools for further research of ovarian cancer.