蚤状蟤(Daphnia pulex) Chk1基因的克隆与表达分析  

Clonning and expression analysis of Chk1 gene in Daphnia pulex

作　　者：方增冰 戴新宇[1] 邹秀[1] 郭晓鸽[1] 徐善良[1] 王丹丽[1] 赵云龙[2] 

机构地区：[1]宁波大学海洋学院,宁波315211 [2]华东师范大学生命科学学院,上海200062

出　　处：《生物学杂志》2015年第2期12-16,57,共6页Journal of Biology

基　　金：国家自然科学基金项目(31172043);浙江省自然科学基金项目(LY12C19003);上海市科技创新行动计划项目(12391900700);宁波市创新团队项目(2011B81003);宁波大学胡岚优秀博士基金

摘　　要：用RACE技术从蚤状溞（ Daphnia pulex）中克隆到Chk1基因cDNA全长为1767 bp，开放阅读框为1497 bp，编码了498个氨基酸，其结构中存在3个保守的Ser-Gln （ SQ）和Thr-Gln （ TQ）序列。同源性比对结果显示，蚤状溞Chk1基因与丽色扇头蜱、切叶蜂、转基因捕食螨、豌豆长管蚜和黑腹果蝇等的同源性均为51％～55％。进化分析发现，蚤状溞Chk1基因与豌豆长管蚜、切叶蜂、黑腹果蝇、丽色扇头蜱和转基因捕食螨等节肢动物亲缘关系最近。Real Time PCR实验结果表明，Chk1 mRNA在两性溞的表达量显著高于孤雌溞（ P＜0.05），且在休眠卵中表达量最低。推测Chk1基因可能在蚤状溞的生殖转化调控中发挥重要作用。The full-length cDNA of a Chk1 （DpChk1） was cloned from cladoceran Daphnia pulex using rapid amplification of comple-mentary DNA ends （ RACE） method.The DpChk1 cDNA is 1767 bp in length;and it has a 1497-bp open reading frame that encodes a 498-amino-acid polypeptide containing three conserved Ser-Gln （SQ）, and Thr-Gln （TQ） sequence.In addition, DpChk1 shared homology of 51%-55% with gene in Rhipicephalus pulchellus, Megachile rotundata, Metaseiulus occidentalis, Acyrthosiphon pisum and Drosophila melanogaster.Phylogenetic analysis revealed that DpChk 1 protein has a close genetic relationship with Phylum arthropo-da such as Acyrthosiphon pisum, Megachile rotundata, Drosophila melanogaster, Rhipicephalus pulchellus, Metaseiulus occidenat lis and so on.qPCR results （real-time quantitative PCR） showed that the DpChk1 expression was significantly higher （P〈0.05） in ephippial female than in parthenogenetic female and was the lowest in the resting egg .Therefore, Chk1 was closely related to the reproduction conversion of Daphnia pulex.

关 键 词：蚤状溞 CHK1 REAL time-PCR 生殖转化 

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