鸡Slc24a5基因的cDNA克隆、表达分析及其与黑色素沉积的关系研究

黑色素是一种由醌/吲哚-醌来源的混合物组成的生物高分子化合物.目前在小鼠中已发现的和黑色素沉积相关的基因已经超过了100个.Slc24a5(solute carrier family 24,member 5)基因是在斑马鱼中克隆的一个新基因.研究表明,Slc24a5基因可以调控斑马鱼中的黑色素沉积.鸡作为一种模式动物,已经广泛地被应用于实验研究.为了研究Slc24a5基因在鸡中的情况,克隆了鸡Slc24a5基因全长CDS,并且分析了其与黑色素沉积的关系.鸡Slc24a5基因全长CDS为1 269 bp,编码一个423氨基酸残基的蛋白质.该蛋白质比哺乳动物中的少大约80个氨基酸残基.基因全长超过11 kb,包含8个内含子和9个外显子.RT-PCR结果显示,鸡Slc24a5基因在多处组织表达(眼、脑、皮、肉、心、肝、肾和肺).通过荧光实时定量PCR对不同鸡种中的Slc24a5基因表达量进行检测,发现其在白莱杭,乌骨鸡和北京油鸡的眼睛中表达量很高.并且在乌骨鸡的皮肤和肌肉中Slc24a5基因也有很高的表达.乌骨鸡眼睛中的Slc24a5基因表达量为白莱杭的2倍.而在乌骨鸡皮肤中,Slc24a5基因表达量为白莱杭的70倍,为北京油鸡的15倍.Slc24a5基因在乌骨鸡肌肉中的表达量为白莱杭的15倍,为北京油鸡的3倍.同时通过对这3个鸡种中黑色素在各组织中沉积量进行分析,发现黑色素沉积越多的地方,Slc24a5基因表达量越高.这些结果表明鸡Slc24a5基因的表达与鸡中黑色素的沉积相关.

cDNA Cloning, Expression Analysis of Slc24a5 and Its Relationship With Melanin Deposition in Chicken

Melanin is a kind of biopolymer, composed of a complex quinine/indole-quinone derived mixture which is produced in melanocytes from tyrosine. More than 100 genes related to melanin deposition have been found in mouse. Slc24a5 (solute carrier family 24 , member 5) is a novel gene first cloned in zebrafish. And it has been confirmed that the Slc24a5 is involved in controlling the melanin deposition in zebrafish. Here the full length of the chicken slc24a5 mRNA sequence, its expression profile and a discussion of its relationship to melanin deposition in chicken were reported. Chicken Slc254a5 has a CDS of 1 269bp, predicting a protein of 423 amino acids which is about 80 amino acids shorter than in human and mouse. It has 9 exons and 8 introns and spans more than 11kb of genome sequence. The quantitative real-time PCR confirmed that the chicken Slc24a5 was highly expressed in the eye of White Leghorn and Beijing Fatty Chickens, and in the eye, skin and muscle of Silky. The relative expression in Silky eye is more than two times that of White Leghorn. The relative expression in Silky skin is about 70 times that of White Leghorn and about 15 times that of Beijing Fatty Chickens. The relative expression in Silky muscle is about 15 times that of White Leghorn and about 3 times that of Beijing Fatty Chickens. And the expression result is in accordance with the melanocyte and melanin deposition in these tissues.