鹌鹑piRNAs和Piwil1基因克隆及表达研究  

Cloning and Expression Analysis of piRNAs and Piwil1 Gene in Quail

作　　者：陈蓉[1] 常国斌[1] 张颖[1] 陈国宏[1] 胡国顺[1] 栾德琴[1] 刘艳[1] 戴爱琴[1] 

机构地区：[1]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《中国农业科学》2011年第8期1727-1735,共9页Scientia Agricultura Sinica

基　　金：国家自然科学基金资助(30700564);江苏省自然科学基金资助(BK2009190)

摘　　要：【目的】目前,国内外尚无有关禽类piRNAs及其结合蛋白Piwi的研究报道,开展禽类小分子RNA及其结合蛋白的研究将为小分子RNA的遗传特性、发生和消失机理以及转基因鸡等研究提供基础依据。【方法】以鹌鹑为研究对象,通过构建cDNA文库和TA克隆测序的方法从睾丸组织中获得piRNAs序列和Piwil1基因的CDS序列,同时采用Q-PCR技术分析了鹌鹑不同生命阶段不同组织中piRNAs和Piwil1基因的表达规律。【结果】从鹌鹑睾丸组织中克隆了13个piRNAs序列和Piwil1基因的CDS序列;Q-PCR结果表明piRNAs和Piwil1基因在鹌鹑不同生命阶段不同组织中均有不同程度地表达。【结论】鹌鹑piRNAs长度与哺乳动物相似;Piwil1基因与红色原鸡有较高的同源性,包含PAZ和Piwi结构域,无信号肽剪切位点,推测其属于Piwi家族蛋白,可能通过降低结合RNA的能力或者影响其它转录因子与结构基因启动子的亲和性发挥调控作用;在睾丸组织中,piRNAs和Piwil1蛋白的表达量均是随着个体的不断发育而增加,推断在禽类中piRNAs可能与Piwi蛋白相互作用共同调控精子的发生过程。【Objcetive】A study on small RNA and its binding protein in poultry was carried out in order to provide a basic foundation for the study of genetic characters,occurrence and disappearance mechanisms of small RNAs in avain and transgenic chicken in avian.【Mehtod】The sequences of piRNAs and CDS of the Piwil1 gene were acquired from the testis of quail by constructing cDNA library and TA cloning and sequencing.Next,the expressions of piRNAs and Piwil1 in quail tissues of different stages of life were analyzed by Q-PCR.【Result】Ithis study,we obtained Thirty piRNAs cloned sequences and CDS of the Piwil1 gene were obtained from the testis of quail for the first time.The results of Q-PCR showed that piRNAs and Piwil1 genes had a variable expression in quail tissues of different stages of life.【Conclusion】The results showed that the length of quail piRNAs was similar to mammals.In quail,Piwil1 gene had high homology with that of the red jungle fowl,whose protein contains the PAZ and Piwi domain,and has no signal peptide splice site,suggesting that it belong to Piwi family and might play a regulatory role by reducing the capacity of binding RNA or affecting the affinity between other transcription factors and the promoter of structural gene.In testis,the expression of piRNAs and Piwil1 gene increased with the development of individuals,so it was inferred that piRNAs might regulate the process of spermatogenesis through interacting with Piwi proteins in poultry.

关 键 词：鹌鹑 睾丸 piRNAs PIWI 克隆 

分 类 号：S839[农业科学—畜牧学]