紫红笛鲷过敏原小清蛋白基因克隆及序列分析

目的 克隆紫红笛鲷(Lutianus argentimaculatus)过敏原小清蛋白(parvalbumin)基因。方法 采用生物信息学方法对多种鱼的小清蛋白基因进行序列性比对,根据序列保守区域设计并合成简并性引物,通过实时(RT)-PCR和3'cDNA末端快速扩增RACE技术克隆紫红笛鲷小清蛋白的全长基因,并进行序列分析。结果 获得紫红笛鲷全长608 bp的新基因,开放阅读框为330个碱基,编码109个氨基酸,经分析该蛋白分子量约为11.5kD,等电点为4.35。序列分析结果显示所克隆的基因与其他鱼类过敏原小清蛋白基因有很高的同源性。结论 成功克隆出紫红笛鲷过敏原小清蛋白基因(登录号为:EF591789)。

Cloning and sequence analysis of allergenic parvalbumin gene from red snapper(Lutianus argentimaculatus)

Objective To clone allergenic parvalbumin genes from red snapper.Methods All the known parvalbumin gene sequences from several fish species were compared by bioinfor matic method and the highly conserved domains were selected for degenerative primer designing.The RT-PCR and 3'-RACE methods were applied to clone the full-length parvalbumin cDNA from red snapper.Results Our data indicated that the full-leng th cDNA of red snapper parvalbumin containsed 608 bp nucleotides.It encodes a 109 amino acid protein with an estimated molecular weight of approx imately 11.5 kD and an isoelectric point of 4.35.The predicted pro tein sequence showed high homology with aller genic parvalbumin from many other fish species.Conclusion An allergenic par valbumin gene was cloned from red snapper(access number: EF591789).