不同倍性鲫鲤MeCP2基因分子克隆及时空表达分析  

Molecular cloning and spatio-temporal expression characteristic analysis of MeCP2 gene in different ploidy cyprinid fishes

作　　者：周蓉[1] 伍艳红[1] 姚潇[1] 凌欣[1] 刘文彬[1] 刘少军[1] 

机构地区：[1]湖南师范大学生命科学学院,湖南长沙410081

出　　处：《水产学报》2016年第1期1-10,共10页Journal of Fisheries of China

基　　金：中国博士后科学基金(2014M560645;2015T80874);国家自然科学基金重点项目(31430088);国家自然科学基金重大国际合作项目(31210103918);湖南省科技计划资助(2014FJ3084;2015RS4031);湖南省生物发育工程及新产品研发协同创新中心(20134486)~~

摘　　要：为了探寻不同倍性鱼生殖育性的表观遗传调控规律,实验基于不同倍性鲫鲤精巢的转录本信息库设计引物克隆了不同倍性鲫鲤的MeCP2基因。序列比对结果发现,在远缘杂交的过程中基因组发生了遗传重组和变异。基于序列公共部分设计引物,通过半定量PCR和实时定量PCR的方法分析了MeCP2基因在不同倍性鱼发育过程中的时空表达特征。结果还发现,MeCP2基因在所有组织中都有表达,但在脑组织中的表达量最高;纵向对比性腺的发育过程,MeCP2基因的表达量会随着性腺发育成熟而下降;横向对比不同倍性鱼的性腺发育,MeCP2基因在三倍体鱼卵巢中的表达量明显高于二倍体和四倍体鱼,但在精巢中的表达量随鱼的倍性增加而增加。研究表明,MeCP2基因的表达与不同倍性鱼的卵巢发育密切相关,这为研究鱼类生殖不育的分子机理及其在水产育种上的应用奠定了坚实的基础。Distant hybridization is the important pathway of forming hybrid vigor and polyploid individuals. By combined distant hybridization and interploid hybridization, we have obtained different ploid crucian carp. MeCP2 is a kind of DNA methylation binding protein and functions as a crucial gene expression regulator by translating epigenetic DNA methylation to proper cell response. To discover the epigenetic regulation mechanism of the reproductive ability in different ploid crucian carp, this study cloned the MeCP2 gene in different ploid crucian carp based on the transcriptome of testis. Sequence alignments indicated that there are genetic recombination and variation in genome during distant hybridization. A pair of common primers was designed to analyze the spatial and temporal expressional characteristics of MeCP2 gene during the development process of different ploid fish through semi-RT PCR and real-time PCR. The results showed that MeCP2 gene was expressed widely with the highest expression in the brain. Compared the different stages of gonadal development, the expression of MeCP2 gene decreased coupled with gonad maturation. While compared the different ploid crucian carp at the same developmental stages, MeCP2 gene was expressed significantly higher in the ovary of triploid fish than in diploid and tetraploid fish, while increased in the testis coupled with the ploid change. Taken together, the expression of MeCP2 gene is closely related to ovary maturation of different ploid fish, which lays the foundation for studying the mechanism and application of fish infertility in genetic breeding and aquaculture.

关 键 词：异源三倍体鲫鲤 异源四倍体鲫鲤 MECP2基因 性腺发育 

分 类 号：Q785[生物学—分子生物学] S965[农业科学—水产养殖]