刺激隐核虫14-3-3基因的分子特征

谢金珠; 单金红; 徐慧源; 黄晓红

0
0
浏览
下载

摘要
关键词
基金信息
论文图表
同行评议
相关论文
评论

刺激隐核虫14-3-3基因的分子特征

首发时间：2016-02-26

谢金珠 ¹
谢金珠（1989-），女，硕士，主要从事病原分子生物学的研究
单金红 ¹ 徐慧源 ¹ 黄晓红 ¹
黄晓红（1963-），女，教授，病原分子生物学等

1、福建师范大学生命科学学院福建省发育与神经生物学重点实验室，福州 350117

摘要：刺激隐核虫病作为海水硬骨鱼的致死性传染病严重制约了海产养殖业的可持续发展，对其病原生物学及防治的研究具有重要的意义。14-3-3蛋白是真核生物体内重要的多功能调控蛋白，对某些寄生虫感染具有一定的免疫保护作用。本实验从刺激隐核虫的滋养体cDNA文库中克隆了一个新的14-3-3蛋白同源基因（Ci14-3-3-Y251）（GenBank KU711805）；其全长cDNA为824bp，开放阅读框(ORF)为732bp，编码一条含243个氨基酸、理论分子量为28kDa的多肽链；该ORF中有7个密码子（6个TAA和1个TAG）是纤毛虫里特有的，将它们改为相应的通用密码子后，人工合成完整的ORF，并将其通过载体pGEX-4T-1亚克隆到大肠杆菌中；以乳糖自诱导法诱导细菌表达Ci14-3-3-Y251，获得大量重组蛋白rCi14-3-3-Y251；以纯化的rCi14-3-3-Y251免疫小鼠获得多克隆抗体，用于刺激隐核虫内源14-3-3-Y251蛋白的免疫印迹分析。结果显示：rCi14-3-3-Y251具有较好的抗原性；内源Ci14-3-3-Y251蛋白的表观分子量为28 kDa，与理论值相符，该蛋白在刺激隐核虫的各个时期均有表达。本实验对刺激隐核虫14-3-3基因功能及应用潜能的研究打下了基础。

关键词：兽医寄生虫学刺激隐核虫 14-3-3蛋白原核表达免疫原性

For information in English, please click here

Molecular characterization of a novel 14-3-3 gene from Cryptocaryon irritans

XIE Jinzhu ¹
谢金珠（1989-），女，硕士，主要从事病原分子生物学的研究
SHAN Jinhong ¹ XU Huiyuan ¹ HUANG Xiaohong ¹
黄晓红（1963-），女，教授，病原分子生物学等

1、Fujian Key Laboratory of Developmental Biology and Neuroscience, College of Life Science, Fujian Normal University, Fuzhou 350117

Abstract：Cryptocaryosis is an infectious disease lethal to seawater teleost fishes, which has hindered the sustainable development of marine aquaculture. Therefore, it is of importance to study the pathogenic biology for the control of cryptocaryosis. 14-3-3 proteins are a family of regulation proteins with multifunction, involved in a lot of critical cellular activities. It was found that vaccines with 14-3-3 protein had induced murine immune protections from several kinds of parasite infections to certain extent. In this experiment, a novel 14-3-3 gene homolog (GenBank KU711805)(Ci14-3-3-Y251) was cloned from the cDNA library of C. irritans trophonts. Its full-length cDNA is 824 bp with an open reading frame (ORF) of 732 bp, which encoded a polypeptide of 243 amino acids. The calculated molecular weight of the polypeptide was 28 kDa. There were 7 non-universal codons (6 TAA and 1 TAG) in the ORF, which was modified to the corresponding universal codons for glutamine for the proper expression in Escherichia coli. The modified ORF was synthesized artificially, amplified, then inserted into BamHI and SalI site of plasmid pGEX-4T-1 and transformed into E. coli. The bacteria with pGEX-4T/Ci14-3-3-Y251 was induced to expression foreign gene by the addition of lactose. As a result, the recombinant Ci14-3-3-Y251 proteins was expressed at high-level. Polyclonal antibody against rCi14-3-3-Y251 was prepared by immunizing the SPF mice with the purified rCi14-3-3-Y251, and used to detect the endogenous Ci14-3-3-Y251 at various developmental stages of C. irritans. The results showed that rCi14-3-3-Y251 possessed good antigenicity, the molecular mass of the endogenous Ci14-3-3-Y251 was correspondent with the calculated one. The result also showed that the endogenous Ci14-3-3-Y51 was expressed at all developmental stages of C. irritans. This experiment would be beneficial for further researches on the control of cryptocaryosis.

Keywords： veterinary parasitology；Cryptocaryon irritans；14-3-3 protein；prokaryotic expression immunogenicity

基金：

论文图表：

引用

导出参考文献

.txt

.ris

.doc

谢金珠，单金红，徐慧源，等. 刺激隐核虫14-3-3基因的分子特征[EB/OL]. 北京：中国科技论文在线 [2016-02-26]. https://www.paper.edu.cn/releasepaper/content/201602-186.

No.4678559106145214****

同行评议

共计0人参与

全部评论

0/1000

论文编号	201602-186
论文题目	刺激隐核虫14-3-3基因的分子特征
文献类型
收录期刊	上传封面中文期刊英文期刊期刊名称（中文）期刊名称（英文）年，卷（）上传封面中文专著英文专著书名（中文）书名（英文）出版地出版社出版年上传封面中文译著英文译著书名（中文）书名（英文）出版地出版社出版年上传封面中文论文集英文论文集编者.论文集名称（中文） [c]. 出版地出版社出版年， - 编者.论文集名称（英文） [c]. 出版地出版社出版年，- 上传封面中文文献英文文献期刊名称（中文）期刊名称（英文）日期-- 在线地址http:// 上传封面中文文献英文文献文题（中文）文题（英文）出版地出版社,出版日期-- 上传封面中文文献英文文献文题（中文）文题（英文）出版地出版社,出版日期--
英文作者写法：中外文作者均姓前名后，姓大写，名的第一个字母大写，姓全称写出，名可只写第一个字母，其后不加实心圆点“.”, 作者之间用逗号“，”分隔，最后为实心圆点“.”, 示例1：原姓名写法：Albert Einstein,编入参考文献时写法：Einstein A. 示例2：原姓名写法：李时珍；编入参考文献时写法：LI S Z. 示例3：YELLAND R L,JONES S C,EASTON K S,et al.