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细粒棘球绦虫锌指蛋白基因的克隆、序列分析及在不同发育阶段的表达分析
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引用本文:徐梦飞,卢平萍,马 勋,张艳艳,王炜烨,孟季蒙,王正荣,薄新文.细粒棘球绦虫锌指蛋白基因的克隆、序列分析及在不同发育阶段的表达分析[J].西北农业学报,2018,27(12):1707~1715
DOI:10.7606/j.issn.1004-1389.2018.12.001
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徐梦飞,卢平萍,马 勋,张艳艳,王炜烨,孟季蒙,王正荣,薄新文 (1.石河子大学 动物科技学院新疆石河子 832000 2.新疆农垦科学院 畜牧兽医研究所省部共建绵羊遗传改良与健康养殖国家重点实验室新疆石河子 832000) 
基金项目:国家重点基础研究发展计划(973计划)(2015CB150300);国家自然科学基金(31360608);新疆生产建设兵团科技攻关项目(2014BA002);新疆生产建设兵团国际科技合作项目(2013BC001)。
中文摘要:旨在分析并预测细粒棘球绦虫锌指蛋白(Echinococcus granulosus zinc finger peotein,Eg-ZFP)结构、功能及其在细粒棘球绦虫发育过程中的表达模式,预测其在细粒棘球绦虫防治中潜在的作用。通过对前期细粒棘球绦虫原头蚴(protoscolex,PSC)转录组高通量测序结果中Unigene进行筛选,克隆Eg-ZFP基因并进行序列分析,实时荧光定量PCR(Realtime fluorescence quantitative PCR,qPCR)分析Eg-ZFP在原头蚴及成虫中的表达特征,利用全量组织原位杂交(Whole mount in situ hybridization,WISH)检测该蛋白mRNA在原头蚴和成虫组织中的分布情况。 结果显示,细粒棘球绦虫Eg-ZFP基因的CDS序列长852 bp,具有完整的开放阅读框(852 bp),编码283个氨基酸,预测表明Eg-ZFP分子质量大小及等电点为31.6 ku和8.89;qPCR检测发现,Eg-ZFP在原头蚴及成虫阶段均有表达,且在成虫阶段表达量高;全量组织原位杂交检测结果表明,Eg-ZFP mRNA在原头蚴及成虫中分布广泛,且在成虫的生殖系统中丰度较高,提示Eg-ZFP与虫体生殖发育有关。结果表明Eg-ZFP在细粒棘球绦虫发育过程中具有重要作用,为进一步研究Eg-ZFP在E.granulosus防治中的作用奠定基础。
中文关键词:细粒棘球绦虫  锌指蛋白  基因克隆  生物信息学分析  实时定量PCR  全量组织原位杂交
 
Cloning,Sequencing,and Expression Analysis at Different Development Stages of Zinc Finger Protein Gene of Echinococcus granulosus
Abstract:The aim of this study is to analyze and predict the structure and function of the Echinococcus granulosus zinc finger protein (Eg-ZFP) and its expression pattern in the different development stages of Echinococcus granulosus (E.g),and predict its potential role in the prevention and control of E.g.Based on the results of the high-throughput sequencing of the protoscolex (PSC) transcriptome,Eg-ZFP gene was successfully cloned and to analyze its sequence characteristics by the bioinformatics software,quantitative real-time PCR (qPCR) was used to confirm the quantity of Eg-ZFP gene expression in different developmental stages of E.g,the distribution feature of Eg-ZFP mRNA was analyzed by whole mount in situ hybridization (WISH) in PSC and adult.The results suggested that the Eg-ZFP of E.g is at the length of 852 bp,containing an open reading frame of 852 bp which encoded 283 amino acids with a predicted molecular mass of 31.6 ku and PI of 8.89.According to the analysis of qPCR,mRNA expression of Eg-ZFP was detected at PSC and adult,the Eg-ZFP mRNA expression levels was higher at adult.WISH detection shows that the Eg-ZFP mRNA is widely distributed in the PSC and adults and have higher abundance in the adult reproductive system,suggesting that Eg-ZFP may be related to the larval reproductive development.These indicated that Eg-ZFP could play an important role in the development of E.g.This will lay a foundation for further research on the role of Eg-ZFP in the prevention and treatment of E.g.
keywords:Echinococcus granulosus  Zinc finger protein  Gene clone  Bioinformatics analysis  Quantitative real-time PCR  Whole mount in situ hybridization
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