刚地弓形虫SRS29C截短型基因的克隆及原核表达质粒的构建

doi:10.19640/j.cnki.jtau.2021.03.011

天津农学院学报 ›› 2021, Vol. 28 ›› Issue (3): 50-55.doi: 10.19640/j.cnki.jtau.2021.03.011

刚地弓形虫SRS29C截短型基因的克隆及原核表达质粒的构建

袁娣¹, 邵怡¹, 张同瑶¹, 姜阜杉¹, 李秀荣², 宋淇淇^{1,通信作者}

1.天津农学院动物科学与动物医学学院天津市农业动物繁育与健康养殖重点实验室,天津 300392;
2.巴彦淖尔市临河区农畜产品质量安全中心,内蒙古自治区巴彦淖尔 015000

收稿日期:2020-04-14 出版日期:2021-09-30 发布日期:2021-10-07
通讯作者: 宋淇淇（1986—）,女,讲师,博士,主要从事兽医寄生虫学教学及研究工作。E-mail：qqs0606@163.com。
作者简介:袁娣（1997—）,女,本科在读,动物医学专业。E-mail：yuandi1217@foxmail.com
基金资助:
天津市自然科学基金项目（19JCQNJC13700）; 天津市教委科研计划项目（2018KJ185）; 天津市大学生创新训练计划项目（201803011）

Cloning and construction of prokaryotic expression plasmid of truncated SRS29C gene of Toxoplasma gondii

Yuan Di¹, Shao Yi¹, Zhang Tongyao¹, Jiang Fushan¹, Li Xiurong², Song Qiqi^{1,Corresponding Author}

1. Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China;
2. Agricultural and Animal Product Quality and Safety Center of Linhe district, Bayannaoer 015000, Inner Mongolia Autonomous Region, China

Received:2020-04-14 Online:2021-09-30 Published:2021-10-07

摘要/Abstract

摘要： 为构建刚地弓形虫SRS29C基因原核表达质粒,利用SignalP-5.0软件预测信号肽序列,去除SRS29C前端信号肽序列,设计引物并扩增SRS29C截短型基因序列,克隆并构建原核表达质粒。结果显示：SRS29C基因前51位氨基酸为信号肽区域,故将此段信号肽序列切除,获得长度为 966 bp的SRS29C截短型基因片段（SRS29Ccut）,成功构建原核表达质粒pET-28a-SRS29Ccut和pET-32a-SRS29Ccut。此研究为今后对刚地弓形虫SRS29C蛋白的生物学功能及免疫特性进行更深入研究打下基础。

关键词: 刚地弓形虫, SRS29C, 截短型基因, 原核表达质粒

Abstract: In order to construct the prokaryotic expression plasmid of Toxoplasma gondii SRS29C gene, the signal peptide of SRS29C gene was predicted by using SignalP-5.0 software, the sequence of the signal peptide of SRS29C gene was removed, the truncated gene of SRS29C was amplified and cloned, and then the prokaryotic expression plasmids were constructed. The results showed that the first 51 amino acids of SRS29C were the signal peptide region, so by cutting off the signal peptide, the 966 bp SRS29C truncated gene fragment (SRS29Ccut) was successfully obtained, and the recombinant plasmids pET-28a-SRS29Ccut and pET-32a-SRS29Ccut were successfully constructed. This study may lay a foundation for further study on the biological function and immune characteristics of the protein of Toxoplasma gondii SRS29C.

Key words: Toxoplasma gondii, SRS29C, truncated gene, prokaryotic expression plasmid

中图分类号:

S855.9

袁娣, 邵怡, 张同瑶, 姜阜杉, 李秀荣, 宋淇淇. 刚地弓形虫SRS29C截短型基因的克隆及原核表达质粒的构建[J]. 天津农学院学报, 2021, 28(3): 50-55.

Yuan Di, Shao Yi, Zhang Tongyao, Jiang Fushan, Li Xiurong, Song Qiqi. Cloning and construction of prokaryotic expression plasmid of truncated SRS29C gene of Toxoplasma gondii[J]. Journal of Tianjin Agricultural University, 2021, 28(3): 50-55.

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刚地弓形虫SRS29C截短型基因的克隆及原核表达质粒的构建

Cloning and construction of prokaryotic expression plasmid of truncated SRS29C gene of Toxoplasma gondii

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