大鳞副泥鳅Sox基因保守区的克隆与特征分析  

Cloning and characterization of conservative region sequences of Sox genes in Paramisgurnus dabryanus

作　　者：游翠红[1,2] 童金苟[1] 俞小牧[1] 

机构地区：[1]中国科学院水生生物研究所/淡水生态与生物技术国家重点实验室,武汉430072 [2]汕头大学海洋生物研究所,汕头515063

出　　处：《华中农业大学学报》2012年第4期490-498,共9页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(30370225);农业部专项课题(200903045;2011-G12)

摘　　要：为探讨大鳞副泥鳅(Paramisgurnus dabryanus)的性别决定与分化机制,采用Sox基因HMG盒保守区的2对简并引物,SoxN与Sox9,用以扩增大鳞副泥鳅的基因组DNA,并对其性腺组织进行RT-PCR分析。SoxN引物扩增基因组DNA共获得pdSox1、pdSox2、pdSox3、pdSox4、pdSox11、pdSox14、pdSox19、pd-Sox21等13个Sox基因,分属SoxB与SoxC类群;Sox9引物扩增基因组DNA共获得pdSox8、pdSox9等5个Sox基因,属于SoxE类群,这5个Sox基因均含有一个内含子,且内含子的起始位置一致。基于Sox基因HMG盒蛋白质的遗传距离构建的NJ系统进化树表明,大鳞副泥鳅18个Sox基因在小鼠或斑马鱼中均能找到相应的Sox直系同源基因,其中,pdSox4、pdSox8、pdSox9、pdSox11、pdSox14、pdSox19等6个基因存在基因加倍现象,具有2个或2个以上的拷贝。用Sox9引物对大鳞副泥鳅的性腺进行RT-PCR扩增和测序分析,结果表明,在精巢中表达的是pdSox9a,在卵巢中表达的是pdSox8a。以上结果表明,pdSox8与pdSox9是大鳞副泥鳅的性别相关基因,它们可能在大鳞副泥鳅的性别分化中发挥作用。In order to further investigate the sex determination and sex differentiation mechanism of large scale loach Paramisgurnus dabryanus, two degenerate primers,SoxN and Sox9, were used to carry out PCR amplification of the genomic DNA and RT-PCR analysis of the gonad tissues. Thirteen genes of the Sox family including pdSox1 , pdSox2, pdSox3, pdSox4, pdSox11, pdSox14, pdSox19 and pdSox21 were cloned from the genomic DNA of P. dabryanus with the SoxN primer and classified as the SoxB and SoxC groups. Another five Sox genes including pdSox8 and pdSox9, with one intron starting at the same position, were obtained by using the Sox9 primer and classified as the SoxE group. Based on the genetic distance of the HMG-box amino acid,all the eighteen pdSox were orthologous to the Sox genes of mouse or zebrafish on the NJ phylogenetic tree. Six of the 13 Sox genes including pdSox4, pdSox8, pdSox9,pdSox11 ,pdSox14 and pdSox19 had two or more copies. Moreover, the results of RT-PCR and sequence analyses with the Sox9 primer showed that pdSox9a was expressed in the testis whereas pdsoxSa was expressed in the ovary. The present results suggested that pdSox8 and pdSox9 were sex-related genes and they might play a role in the sex differentiation of P. dabryanus.

关 键 词：大鳞副泥鳅 SOX基因 HMG盒 性别决定 性别分化 

分 类 号：S917.4[农业科学—水产科学]