论文部分内容阅读
采用荧光定量PCR技术检测了Dmrt2基因的两个亚型Dmrt2a和Dmrt2b在金鱼(Carassius auratus)不同发育时期以及不同组织中的表达情况,用RACE技术克隆得到金鱼Dmrt2a cDNA序列的全长为1 755 bp,5’和3’非编码区长分别为188 bp和67 bp,推测的开放阅读框可编码499个氨基酸的多肽。分子系统进化分析表明金鱼Dmrt2a与其它鱼类Dmrt2a基因聚成一支,与斑马鱼(Daniorerio)、红鳍东方鲀(Takifugu rubripes)、青鳉(Oryzias latipes)、罗非鱼(Oreochromis Niloticus)Dmrt2a的同源性分别为85%、61%、58%、58%。荧光定量PCR结果揭示,Dmrt2a在精巢中表达量最高,在卵巢中次之,而Dmrt2b在卵巢中表达量最高,在精巢中次之,二者在肾脏、消化道、肝脏、心脏和脑中都有表达,但表达量低;不同发育时期胚胎和仔鱼的Dmrt2a和Dmrt2b表达量变化很大,Dmrt2a在受精后24 h达到最高值,之后表达量降低,而Dmrt2b在孵化后仔鱼中表达量显著增加。
Fluorescent quantitative PCR was used to detect the expression of Dmrt2a and Dmrt2b of Dmrt2 gene at different developmental stages and different tissues of goldfish (Carassius auratus). The full-length cDNA sequence of Dmrt2a was 1 755 bp The 5 ’and 3’ noncoding regions were 188 bp and 67 bp, respectively. The deduced open reading frame (ORF) encoded 499 amino acids. Molecular phylogenetic analysis showed that the goldfish Dmrt2a was clustered with other fish Dmrt2a genes and was found to associate with Daniorerio, Takifugu rubripes, Oryzias latipes, and Oreochromis niloticus Dmrt2a Source were 85%, 61%, 58%, 58%. Quantitative real-time PCR revealed that Dmrt2a was the highest expression in the testis and secondarily in the ovary, whereas Dmrt2b was the highest in the ovary and secondarily in the testis, both in the kidney, digestive tract, liver, heart and brain Dmrt2a and Dmrt2b in embryos and larvae at different developmental stages varied greatly. Dmrt2a reached the maximum at 24 h after fertilization, and then decreased, while the expression of Dmrt2b was significantly increased in larvae after incubation .