水产学报2017，Vol.41Issue(7)：1017-1027,11.DOI:10.11964/jfc.20161210654
一种鰑鱼诺卡氏菌酪氨酸蛋白磷酸酶基因的克隆及功能初步研究
Gene cloning and preliminary function study of protein tyrosine phosphatase from Nocardia seriolae
摘要
Abstract
Nocardia seriolae is the main pathogen of fish nocardiosis, which is a chronic systemic granuloma disease of fish. A gene of protein tyrosine phosphatase (PTP) was found by analyzing the whole genome sequence of N. seriolae, and bioinformatics analysis showed that the PTP gene mayencode a secreted protein which could possibly target host cell mitochondria. In this study, the gene cloning, subcellular localization, over-expression and mitochondrial membrane potential detection were carried out. The results showed that the protein PTP was identified in the extracellular products of N. seriolae, which confirmed that PTP was a secreted protein. Subcellular localization of PTP-GFP fusion proteins were evenly distributed in the whole cell of FHM cells, and did not coincide with the distribution of mitochondria, which indicated that the protein PTP was not targeted at mitochondria. Typical apoptotic features, such as nuclear pyrosis and apoptotic bodies, were found when PTP protein was expressed in FHM cells by both subcellular localization and over-expression studies. The mitochondrial membrane potential was significantly damaged in FHM at 48h after the transfection of pc DNA-PTP, which confirmed that PTP was a bacterial protein which can likely induce cell apoptosis. The gene cloning and preliminary function study of PTP fromN. seriolae have laid the foundation for further research on the gene and for promoting the understanding of the pathogenic mechanism of N. seriolae.关键词
這鱼诺卡氏菌/酪氨酸蛋白磷酸酶/分泌蛋白/亚细胞定位/细胞凋亡Key words
Nocardia seriolae/protein tyrosine phosphatase (PTP)/secreted protein/subcellular localization/cell apoptosis分类
生物科学引用本文复制引用
夏立群,童邦卓,徐亮,苏泽杰,陈锐敏,廖保山,鲁义善..一种鰑鱼诺卡氏菌酪氨酸蛋白磷酸酶基因的克隆及功能初步研究[J].水产学报,2017,41(7):1017-1027,11.基金项目
广东省科技发展专项资金(2016A050502061) （2016A050502061）
广东省自然科学基金(2014A030313602) （2014A030313602）
深圳市科技计划项目(JCYJ20170306161613251) （JCYJ20170306161613251）
深圳大鹏新区产业发展专项(KY20160207) （KY20160207）
湛江市科技攻关计划(2014b01043) （2014b01043）
广东省攀登计划(pdjh2016b0234)International S&T Cooperation Projects of Guangdong Province (2016A050502061) （pdjh2016b0234）
Guangdong Natural Science Foundation (2014A030313602) （2014A030313602）
Shenzhen Science and Technology Program (JCYJ20170306161613251) （JCYJ20170306161613251）
Shenzhen Dapeng New District Special Fund for Industry Development (KY20160207) （KY20160207）
Zhanjiang science and technology Program (2014b01043) （2014b01043）
Guangdong Province Climb Plan (pdjh2016b0234) （pdjh2016b0234）
