许氏平鮋( Sebastes schlegeli) CYP19B 基因cDNA克隆及在生殖周期中的表达分析

从许氏平鲉脑组织中克隆到细胞色素P450芳香化酶(P450aromB)(CYP19B)基因核心功能区cDNA。结果表明,许氏平鲉CYP19B主要功能区片段编码369个氨基酸,包括I-螺旋区、Ozol’s肽区及部分芳香化酶特异性保守区。RT-PCR分析表明,CYP19B在雌鱼和雄鱼各组织中表达情况不同,在雌鱼中表达较丰富,但是两者都在性腺和脑中有表达,且脑中的表达量高于性腺。生殖季节表达结果表明,CYP19B在繁殖前期表达较为丰富,退化期未见有表达。CYP19B在精巢处于Ⅲ期的许氏平鲉脑组织中表达最为丰富,在精巢发育Ⅳ期的脑组织表达量最低。上述研究结果说明,CYP19B在雌性和雄性许氏平鲉繁殖周期中均发挥重要生理作用。

CLONGING AND THE EXPRESSION ANALYSIS OF CYP19B GENE cDNA DURING THE REPRODUCTIVE CYCLE OF SEBASTES SCHLEGELI

Using nested RT-PCR technology,we cloned brain cytochrome P450 aromatase(P450aromB) gene(CYP19B) cDNA fragment of the major functional areas of black rockfish(Sebastes schlegeli).The mRNA expression was examined in the gonads of male and female black rockfish(S.schlegeli) at different stages of gonadal development.The cloned cDNA function sequence with an open reading frame of 1109 nucleotides encoded 495 predicted amino acid residues,cross-domain gene family characteristics of the three regions,including the I-helix,Ozol’s peptide region,and some aromatic region.In addition,the sites of I133,E302,P308,D309,T310,R435,C437 which consistent with human aromatase gene play an important role on the activity.RT-PCR analysis showed that the CYP19B was expressed in gonads and brain of male and female,but it was more abundant in the brain than in the gonads.The results showed that periodic expression of CYP19B in the early reproductive gonads is more abundant,and there is no expression at stageⅥ.In the stage Ⅲ,the relative expression level of CYP19B in brain is the highest.In the stageⅣ,the relative expression level was the lowest.The study indicates that,CYP19B may play an important physiological role in both female and male black rockfish reproductive.