猪带绦虫丝氨酸蛋白酶B6的鉴定、表达及抗原性分析

Guang-xue Liu, Shao-hua Zhang, Ai-jiang Guo, Jun-ling Hou, Yan-ling Wei, Shuai Wang, Xue-nong Luo
{"title":"猪带绦虫丝氨酸蛋白酶B6的鉴定、表达及抗原性分析","authors":"Guang-xue Liu,&nbsp;Shao-hua Zhang,&nbsp;Ai-jiang Guo,&nbsp;Jun-ling Hou,&nbsp;Yan-ling Wei,&nbsp;Shuai Wang,&nbsp;Xue-nong Luo","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To identify and express serpin B6 of Taenia solium (Tsserpin B6) and explore its possible use as a diagnostic antigen.</p><p><strong>Methods: </strong>Primers for Tsserpin B6 were designed according to T. solium genome and transcriptome data. The Tsserpin B6 gene was amplified from the total RNA of T. solium cysticercus and subsequently analyzed by bioinformatics. Multiple amino acid sequence alignments of Tsserpin B6 and other parasites serpins were created using the Clustal X1.83. Phylogenetic analyses were performed using the MEGA 6.0. The recombinant expression vector pET-30a-Tsserpin B6 was constructed and expressed in E. coli strain BL21 (DE3). The expressed proteins were purified, isolated by SDS-PAGE, and analyzed by Western blotting using pig serum infected with T. solium cysticerci.</p><p><strong>Results: </strong>The complete reading frame of Tsserpin B6 was 1 131 bp and encoded a protein of 376 amino acids. The encoded protein had a conservative reactive center loop and distinctive domains of NEEGAE and FTVDHPFLF, and harbored 9 potential linear B cell epitopes. The expressed products of Tsserpin B6 mainly existed as an inclusion body, and reacted with pig serum infected with T. solium, resulting in a specific band at the Mr 53 000.</p><p><strong>Conclusion: </strong>The Tsserpin B6 gene was successfully cloned, and its expressed products can be recognized by pig serum infected with T. solium.</p>","PeriodicalId":23981,"journal":{"name":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","volume":"34 4","pages":"334-8"},"PeriodicalIF":0.0000,"publicationDate":"2016-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Identification, Expression and Antigenicity Analysis of Serpin B6 of Taenia solium].\",\"authors\":\"Guang-xue Liu,&nbsp;Shao-hua Zhang,&nbsp;Ai-jiang Guo,&nbsp;Jun-ling Hou,&nbsp;Yan-ling Wei,&nbsp;Shuai Wang,&nbsp;Xue-nong Luo\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To identify and express serpin B6 of Taenia solium (Tsserpin B6) and explore its possible use as a diagnostic antigen.</p><p><strong>Methods: </strong>Primers for Tsserpin B6 were designed according to T. solium genome and transcriptome data. The Tsserpin B6 gene was amplified from the total RNA of T. solium cysticercus and subsequently analyzed by bioinformatics. Multiple amino acid sequence alignments of Tsserpin B6 and other parasites serpins were created using the Clustal X1.83. Phylogenetic analyses were performed using the MEGA 6.0. The recombinant expression vector pET-30a-Tsserpin B6 was constructed and expressed in E. coli strain BL21 (DE3). The expressed proteins were purified, isolated by SDS-PAGE, and analyzed by Western blotting using pig serum infected with T. solium cysticerci.</p><p><strong>Results: </strong>The complete reading frame of Tsserpin B6 was 1 131 bp and encoded a protein of 376 amino acids. The encoded protein had a conservative reactive center loop and distinctive domains of NEEGAE and FTVDHPFLF, and harbored 9 potential linear B cell epitopes. The expressed products of Tsserpin B6 mainly existed as an inclusion body, and reacted with pig serum infected with T. solium, resulting in a specific band at the Mr 53 000.</p><p><strong>Conclusion: </strong>The Tsserpin B6 gene was successfully cloned, and its expressed products can be recognized by pig serum infected with T. solium.</p>\",\"PeriodicalId\":23981,\"journal\":{\"name\":\"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\",\"volume\":\"34 4\",\"pages\":\"334-8\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

目的:鉴定和表达猪带绦虫丝氨酸蛋白酶B6 (Tsserpin B6),并探讨其作为诊断抗原的可能性。方法:根据猪绦虫基因组和转录组数据设计Tsserpin B6引物。从猪尾蚴囊尾蚴总RNA中扩增出Tsserpin B6基因,并进行生物信息学分析。利用clstal X1.83对Tsserpin B6和其他寄生虫蛇形蛋白进行了多个氨基酸序列比对。使用MEGA 6.0进行系统发育分析。构建了重组表达载体pET-30a-Tsserpin B6,并在大肠杆菌BL21 (DE3)中表达。对表达蛋白进行纯化,SDS-PAGE分离,用猪囊虫感染血清进行Western blotting分析。结果:Tsserpin B6全长1 131 bp,编码376个氨基酸的蛋白。该编码蛋白具有保守的反应性中心环和独特的NEEGAE和FTVDHPFLF结构域,并含有9个潜在的线性B细胞表位。Tsserpin B6的表达产物主要以包涵体的形式存在,与猪感染猪螺旋体的血清发生反应,在Mr 53000处产生特异性条带。结论:成功克隆了Tsserpin B6基因,其表达产物可被猪猪螺旋体感染血清识别。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Identification, Expression and Antigenicity Analysis of Serpin B6 of Taenia solium].

Objective: To identify and express serpin B6 of Taenia solium (Tsserpin B6) and explore its possible use as a diagnostic antigen.

Methods: Primers for Tsserpin B6 were designed according to T. solium genome and transcriptome data. The Tsserpin B6 gene was amplified from the total RNA of T. solium cysticercus and subsequently analyzed by bioinformatics. Multiple amino acid sequence alignments of Tsserpin B6 and other parasites serpins were created using the Clustal X1.83. Phylogenetic analyses were performed using the MEGA 6.0. The recombinant expression vector pET-30a-Tsserpin B6 was constructed and expressed in E. coli strain BL21 (DE3). The expressed proteins were purified, isolated by SDS-PAGE, and analyzed by Western blotting using pig serum infected with T. solium cysticerci.

Results: The complete reading frame of Tsserpin B6 was 1 131 bp and encoded a protein of 376 amino acids. The encoded protein had a conservative reactive center loop and distinctive domains of NEEGAE and FTVDHPFLF, and harbored 9 potential linear B cell epitopes. The expressed products of Tsserpin B6 mainly existed as an inclusion body, and reacted with pig serum infected with T. solium, resulting in a specific band at the Mr 53 000.

Conclusion: The Tsserpin B6 gene was successfully cloned, and its expressed products can be recognized by pig serum infected with T. solium.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信