蚤状溞doublesex的克隆及在不同生殖状态时的表达特征  

Cloning and expression features of a Doublesex gene in Daphnia pulex during different reproduction modes

作　　者：王琪[1] 宁俊佳 张清科 张萌萌[1] 周伟[1] 吴仓仓 王丹丽[1] 

机构地区：[1]宁波大学海洋学院,宁波315211

出　　处：《生物学杂志》2015年第5期25-29,47,共6页Journal of Biology

基　　金：国家自然科学基金资助项目(31172043);浙江省自然科学基金资助项目(LY12C19003);宁波市创新团队项目(2011B81003);王宽诚教育基金会资助

摘　　要：利用RACE技术从蚤状溞(Daphnia pluex)中克隆到doublesex(dpdsx),dpdsx c DNA全长为1636 bp,其中开放阅读框(ORF)1008 bp,编码335个氨基酸,含有1个DM结构域和1个Dimersation domain。同源性比对结果显示蚤状溞dpdsx与大型溞(Daphnia magna)同源性最高为77%,其次盔形溞(Daphnia galeata)为61%,模糊网纹溞(Ceriodaphnia dubia)为54%;进化分析发现蚤状溞dpdsx与盔形溞、大型溞、模糊网纹溞等溞科的亲缘关系最近;Real time-PCR实验结果表明,dpdsx在雄溞表达水平最高,其次是孤雌溞(带夏卵)和两性溞(带冬卵),雌溞与雄溞有显著性差异(P>0.05)。表明doublesex有可能在蚤状溞生殖转化和性别分化调控上起一定作用。The full-length cDNA of a doublesex （ dpdsx） gene was cloned from the cladoceran Daphnia pulex using rapid amplification of complementary DNA ends （ RACE ） method.The dpdsx cDNA is 1636 bp in length; and it has a 1008-bp open reading frame （ ORF） that encodes a 335-amino-acid polypeptide containing a DM structural domain and a dimersation domain.Homology comparison showed Daphnia pulex′s dpdsx sharing homology of up to 77%with Daphnia magna, 61% with Daphnia magna and 54% with Ceri-odaphnia dubia.Phylogenetic analysis revealed that Daphnia pulex′s dpdsx had a close genetic relationship with Daphniidae such as D. magna, D.magna, C.dubia and so on.Real time-PCR showed that dpdsx had the highest expression level in the male, followed by the parthenogenetic female （ with summer egg） and sexual female （ with winter egg） .The expression had significant difference （ P〉0.05） between the male and the female.The experiment indicated that doublesex gene may be play a role in reproduction switching and controlling sex differentiation of D.pulex.

关 键 词：蚤状溞 DOUBLESEX REAL time—PCR 生殖转化 

分 类 号：Q78[生物学—分子生物学] S917.4[农业科学—水产科学]