鸡基质蛋白3核定位信号的预测与鉴定

doi:10.11843/j.issn.0366-6964.2018.11.021

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (11): 2486-2495.doi: 10.11843/j.issn.0366-6964.2018.11.021

鸡基质蛋白3核定位信号的预测与鉴定

邓珊珊¹, 高洪波^1,2, 袁超^1,2, 赵佳福^1,2, 倪萌萌^1,2, 段志强^1,2*, 嵇辛勤^1,2*

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州大学高原山地动物遗传育种与繁殖教育部重点实验室, 贵阳 550025

收稿日期:2018-04-26 出版日期:2018-11-23 发布日期:2018-11-23
通讯作者: 段志强,副教授,E-mail:zqduan@gzu.edu.cn;嵇辛勤,教授,E-mail:jxq972@aliyun.com
作者简介:邓珊珊(1991-),女,贵州绥阳人,硕士生,主要从事家禽疫病发病机制研究,E-mail:1176218223@qq.com
基金资助:
国家自然科学基金（31760732；31502074）；贵州省科技计划项目（黔科合平台人才[2017]5788号）；贵州省普通本科高等学校科技成果转化与产业化项目（黔教合KY字[2017]055号）；贵州大学引进人才科研项目（贵大人基合字[2014]10号）

Prediction and Identification of Nuclear Localization Signal of Chicken Matrin 3

DENG Shan-shan¹, GAO Hong-bo^1,2, YUAN Chao^1,2, ZHAO Jia-fu^1,2, NI Meng-meng^1,2, DUAN Zhi-qiang^1,2*, JI Xin-qin^1,2*

1. College of Animal Sciences, Guizhou University, Guiyang 550025, China;
2. Key Laboratory of Animal Genetics, Breeding and Reproduction in Plateau Mountainous Region of Ministry of Education, Guizhou University, Guiyang 550025, China

Received:2018-04-26 Online:2018-11-23 Published:2018-11-23

摘要/Abstract

摘要：

本研究旨在预测和鉴定鸡基质蛋白3（matrin 3，MATR3）的核定位信号（nuclear localization signal，NLS）。通过核定位信号在线预测软件预测鸡MATR3蛋白中存在的假定NLS （putative NLS，pNLS），根据预测结果构建鸡MATR3蛋白pNLS缺失的重组真核表达载体，转染细胞后通过观察缺失体重组蛋白的亚细胞定位来确定其NLS。根据鸡MATR3蛋白NLS邻近碱性氨基酸序列特征，构建NLS邻近氨基酸缺失的重组真核表达载体，转染细胞后通过荧光观察和亚细胞定位分析来确定NLS邻近碱性氨基酸是否参与鸡MATR3蛋白的细胞核定位。另外，对鸡MATR3蛋白NLS在不同物种间的保守性以及对外源蛋白的核输入能力进行分析和检测。结果表明，鸡MATR3蛋白中存在4个pNLS，其中pNLS2（⁵⁹⁶PSDKKSK⁶⁰²）缺失导致鸡MATR3蛋白丧失细胞核定位特征。将构建的鸡MATR3蛋白NLS邻近碱性氨基酸缺失的重组真核表达载体转染细胞，通过荧光观察和亚细胞定位分析，发现鸡MATR3蛋白NLS邻近的碱性氨基酸不参与NLS介导的MATR3蛋白细胞核定位。此外，笔者发现该NLS基序在不同物种的MATR3蛋白中均保守，并且与SV40大T抗原NLS具有相同的核输入能力。鸡MATR3蛋白中存在一个高度保守的NLS （⁵⁹⁶PSDKKSK⁶⁰²），其邻近的碱性氨基酸位点不参与MATR3蛋白的细胞核定位。

Abstract:

This study aimed to predict and identify the nuclear localization signal (NLS) of chicken matrin 3 (MATR3). The putative NLS (pNLS) in chicken MATR3 was predicted by the online NLS prediction software. The recombinant plasmids of chicken MATR3 with the pNLS deletion were constructed and then transfected into cells. The subcellular localization of the pNLS deletants was observed and analyzed to determine the NLS of chicken MATR3. The recombinant plasmids expressing the deletion of basic amino acids close to NLS were also constructed to examine their roles in the nuclear localization of chicken MATR3. In addition, the conservation analysis and nuclear import ability of chicken MATR3 NLS were investigated. The results showed that there were four pNLS existed in chicken MATR3, and the deletion of pNLS2 ⁽⁵⁹⁶PSDKKSK⁶⁰²) disrupted the nuclear localization of chicken MATR3. In addition, the constructed recombinant plasmids expressing the deletion of basic amino acids close to NLS were transfected into cells, and the subcellular localization of the recombinant proteins indicated that the adjacent basic amino acids of NLS did not participate in the NLS-mediated nuclear localization of chicken MATR3. Moreover, we also found that the NLS motif in MATR3 protein were conserved among different species, and also had the same nuclear import ability as the NLS of SV40 large T antigen. One highly conservative NLS (⁵⁹⁶PSDKSK⁶⁰²) is present in chicken MATR3, and its adjacent basic amino acids do not participate in the nuclear localization of MATR3 protein.

中图分类号:

S831
Q71

邓珊珊, 高洪波, 袁超, 赵佳福, 倪萌萌, 段志强, 嵇辛勤. 鸡基质蛋白3核定位信号的预测与鉴定[J]. 畜牧兽医学报, 2018, 49(11): 2486-2495.

DENG Shan-shan, GAO Hong-bo, YUAN Chao, ZHAO Jia-fu, NI Meng-meng, DUAN Zhi-qiang, JI Xin-qin. Prediction and Identification of Nuclear Localization Signal of Chicken Matrin 3[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2486-2495.

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鸡基质蛋白3核定位信号的预测与鉴定

Prediction and Identification of Nuclear Localization Signal of Chicken Matrin 3

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