红鳍笛鲷(Lutjnaus erythropterus)酪氨酸酶相关蛋白1基因克隆及表达分析

MOLECULAR CLONING AND EXPRESSION OF TYRP1 GENE IN LUTJANUS ERYTHEROPTERUS

本文利用RACE技术克隆获得红鳍笛鲷(Lutjnaus erythropterus)酪氨酸酶相关蛋白1基因全长,并利用生物信息学方法对该基因的m RNA序列特征、氨基酸序列特征、分类及系统发生进行了分析。结果表明,在红鳍笛鲷中,酪氨酸酶相关蛋白1基因含有TYRP1a和TYRP1b两个拷贝,其中TYRP1a序列全长3178bp,开放阅读框(ORF)长1566bp,5?端235bp,3?端1377bp;TYRP1b基因c DNA全长1871bp,ORF区长1656bp,5?端89bp,3?端126bp。序列分析发现,TYRP1a和TYRP1b基因与其它物种的同源基因保守性较高,尤其是酪氨酸酶基因家族典型的酶活性位点序列在脊椎动物中具有高度保守性。进化分析发现,TYRP1a和TYRP1b基因的种间同源性高于两个基因间的同源性。荧光定量PCR数据分析表明,TYRP1a和TYRP1b基因都在眼睛部位具有最高表达,TYRP1b在黑色皮肤也有较高表达。结果可为进一步阐述TYRP1基因在红鳍笛鲷身体和眼睛部位的色素合成机制提供一定的理论基础。

To explore the gene repertory of TYRP1 of Lutjanus erythropterus, we cloned the full length of two TYRP1（TYRP1a and TYRP1b） genes by using rapid-amplification of c DNA ends（RACE） technology based on the unigene sequence annotated as TYRP1 in the transcriptome database of L. erythropterus. The full lengths are 3178 bp and 1871 bp, encoding 521 and 551 amino acids, respectively. Multiple sequence alignment showed both TYRP1 genes were highly conserved in vertebrate. We found that both genes contained a typical domain of tyrosinase family as revealed with the SMART software. Phylogenetic analysis showed that fish TYRP1 duplicates might have shared an ancient whole genome duplication（WGD） event in the common ancestor. In addition, real-time PCR indicated that both genes could be found in the eyes; however, divergence expression was also found between the two genes, apart from the eyes; and TYRP1 b was also found in the skin. The findings may offer a theoretical basis for elucidating mechanism of pigment formation in the body and the eyes of the fish.