摘要
HP基因编码蛋白是本实验室近期发现的一种鸡柔嫩艾美耳球虫的外分泌蛋白。本试验旨在构建重组荧光真核表达质粒pEGFP-HP,将其转染至细胞内,以此来分析HP基因对宿主细胞凋亡的影响。以柔嫩艾美耳球虫第2代裂殖子cDNA为模板进行PCR扩增,然后克隆至荧光真核表达质粒pEGFP,构建重组荧光真核表达质粒pEGFP-HP,将其转染至鸡巨噬细胞(HD11),42 h后采用流式细胞术检测宿主细胞凋亡情况。菌液PCR鉴定和测序鉴定结果显示,重组荧光真核表达载体pEGFP-HP构建成功。流式细胞术检测发现,pEGFP-HP重组质粒组的早期凋亡率显著高于pEGFP空质粒组和空白对照组(P<0.01)。结果表明柔嫩艾美耳球虫HP基因会促进HD11的细胞凋亡,尤其是早期细胞凋亡。本试验为进一步研究HP基因的生物学功能及其作用机制提供了依据,且将HP基因从此命名为柔嫩艾美耳球虫凋亡诱导基因(IA)。
Eimeria tenella HP protein is an excretory protein recently identified by our laboratory. This study examined the effect of HP gene coding sequence on apoptosis of chicken macrophage HD11 cells through construction of the recombinant fluorescent eukaryotic expression plasmid pEGFP-HP, and subsequent transfection and assessment. This involved PCR amplification of HP coding sequence using the cDNA of second generation merozoites of E.tenella as a template, cloning of the amplified product into fluorescent eukaryotic expression vector pEGFP, transfection of recombinant plasmid pEGFP-HP into HD11 cells, and assessment of cell apoptosis using flow cytometry 42 h after transfected. The results indicated that the rate of early apoptosis in HD11 cells containing recombinant plasmid increased significantly(P<0.01) in comparison with HD11 cells containing vector only or HD11 cells without vector. Given its ability to induce apoptosis of HD11, especially early apoptosis, E.tenella HP gene is renamed E.tenella apoptosis induction(IA) gene. Further study on the biological function and regulating mechanisms of E.tenella IA gene is clearly warranted.
作者
许中衎
王黎霞
张榕珍
张一弛
芳卉
潘晨帆
张建军
安健
XU Zhong-kan;WANG Li-xia;ZHANG Rong-zhen;ZHANG Yi-chi;FANG Hui;PAN Chen-fan;ZHANG Jian-jun;AN Jian(College of Animal Science and Technology,Beijing University of Agriculture,National Animal Experimental Demonstration Center,Beijing 102206,China;Department of Animal Science and Veterinary Medicine,Beijing Vocational College of Agriculture,Beijing 102442,China)
出处
《中国兽医杂志》
CAS
北大核心
2021年第6期34-38,42,共6页
Chinese Journal of Veterinary Medicine
基金
北京市特色高水平骨干专业群项目-动物医学专业群项目-技术平台与社会服务建设项目(PMX2020-157102-00060-15)
北京农业职业学院院级科研项目(XY-JT-03)。
