摘要
利用DD RT-PCR技术筛选出牡蛎中受雌激素调节的基因并对其进行测序,得到5条有注释的基因。采用RACE的方法获得了P450基因1400bp的cDNA序列,其中包含一个975bp的开放阅读框。序列比对表明,该基因属于CYP4V2家族。系统进化树分析表明所获得的P450序列与袖扣海兔螺的CYP4V1和CYP4V2并到一起,与其他无脊椎动物的P450基因具有共同起源,但与脊椎动物的P450基因起源不同。应用Real-time PCR技术检测雌、雄长牡蛎在雌激素刺激后不同时间段P450的转录水平的变化。结果表明在17β-雌二醇刺激后,雌性牡蛎处理组性腺组织P450表现出具有显著的时间依赖性,在性腺被刺激后5d和10d时,P450基因的表达量达到最高,而雌激素对雄性牡蛎性腺组织P450的表达无显著影响。雌激素可能通过对P450的表达正反馈调控影响雌性牡蛎的性腺发育。
Estrogen responsive genes were screened by DD RT-PCR in development of Pacific oyster（Crassostrea gigas）gonads.Five genes with annotation in the GenBank were obtained including the P450-like gene.A gene fragment of 1400 bp with an open reading frame of 975 bp was cloned by RACE.Sequence alignments suggested that this gene belong to the CYP4V2 subfamily.Phylogenic analysis showed that this gene was clustered with the CYP4V1 and CYP4V2 in Cyphoma gibbosum and originated from the same ancestor as other invertebrates but from different ancestor with vertebrates.Expression of this gene was up-regulated by treatment with estrogen in the female oyster gonads but not in the male gonads.Estrogen stimulated the later gonadal development in the female Pacific oyster gonads through a positive feedback system involving P450.
出处
《水产科学》
CAS
北大核心
2012年第6期339-345,共7页
Fisheries Science
基金
山东省自然科学基金资助项目(2007ZRB01905)
