畜牧与兽医2018，Vol.50Issue(1)：54-58,5.
真核表达质粒pcDNA.3.1(+)-ACE2的构建及其在CHO细胞中的表达
Construction of eukaryotic expression plasmid pcDNA3.1 (+)-ACE2 and its expressions in CHO cells
摘要
Abstract
To construct eukaryotic expression plasmid pcDNA3.1 (+)-ACE2 and observe its expression in CHO,total RNA was extracted from kidney of goat and then ACE2 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR).The product was cloned into pcDNA3.1 (+) vector,and they were identified by PCR and double restrictive edonuclease digestion and sequence analysis.Then the recombinant expression plasmid pcDNA3.1 (+)-ACE2 was transfected into CHO cells by lipofectamine 3 000,and the ACE2 protein expression was identified by Western blot.The results showed that RT-PCR product was around 2 200 bp.Analysis by restricting enzyme digestion and DNA sequencing showed the pcDNA3.1 (+)-ACE2 eukaryotic expression plasmid was successfully established,and the expression of pcDNA3.1 (+)-ACE2 could be detected in CHO cells by western blotting.The pcDNA3.1-CSRP2-HA eukaryotic expression plasmid was constructed successfully,and it could be expressed in CHO cells.This study lays the foundation for the research of the activity and immunological functions of ACE2 protein.关键词
血管紧张素转化酶2 (ACE 2)/真核表达质粒/转染/CHO细胞Key words
angiotensin angiotensin converting enzyme 2 (ACE 2)/eukaryotic expression plasmid/transfection/CHO cells分类
农业科技引用本文复制引用
肖航,王凯,王换换,王鲲,张源淑..真核表达质粒pcDNA.3.1(+)-ACE2的构建及其在CHO细胞中的表达[J].畜牧与兽医,2018,50(1):54-58,5.基金项目
国家自然科学基金(30871838) （30871838）
