期刊论文

中文

中国兽医科学

1673-4696

2022

52

1

111-119

10.16656/j.issn.1673-4696.2022.0022

国家自然科学基金青年基金项目（31802151）； 湖南省自然科学基金青年基金项目（2019JJ50257）； 中国博士后科学基金面上项目（2018M642981）； 湖南省教育厅优秀青年项目（18B099）；

本校为第一机构

为了研究胞质视黄酸结合蛋白2(cytoplasmretinoic acid binding proteins 2,Crabp2)对猪卵泡发育的调控作用及机制,笔者通过构建猪Crabp2过表达载体并设计Crabp2 siRNA干扰片段,揭示它们对猪卵泡颗粒细胞凋亡的影响。通过从屠宰场回收猪卵巢,应用TRIzol法提取组织总RNA,并反转录成cDNA。设计Crabp2基因全长引物,利用PCR克隆Crabp2基因,构建Crabp2过表达载体。同时,设计并筛选出干扰效率最高的Crabp2 siRNA。将Crabp2过表达载体和siRNA片段分别转染至猪卵泡颗粒细胞中24 h,利用实时荧光定量PCR(qRT- PCR)和蛋白质免疫印迹(Western- blot)检测颗粒细胞Crabp2的表达水平,流式细胞术检测细胞凋亡率的...

In order to reveal the regulation effect and mechanism of cytoplasmic retinoic acid binding protein 2(Crabp2) on porcine follicular development,Crabp2 overexpression vector was constructed and Crabp2 si RNA was designed to investigate their effects on apoptosis in porcine follicular granulosa cells.Total RNA was extracted from pig ovary from slaughterhouses and reverse transcribed into c DNA in this study.The specific primer of Crabp2 gene was designed.The Crabp2 gene was cloned by PCR,and Crabp2 overexpression plasmid was constructed successfully.Meanwhile,Crabp2 si RNA with the highest inter...