野猪RBP4基因的克隆、表达及变异初步研究

Cloning,expression and mutations analysis of RBP4 gene of wild boar

视黄醇结合蛋白是体内一类将维生素A从肝中转运至靶组织以实现维生素A的细胞内转运代谢的特异的运载蛋白,在协助维生素A发挥生理功能中起着不可替代的作用。它既是一个影响猪的繁殖性状的候选基因,同时也是一个研究维生素A代谢障碍及肥胖症的候选基因。为了进一步揭示其群体遗传变异,寻找新的遗传标记,本研究对野猪(Sus scrofa ussuricus)RBP4基因进行了克隆、表达和序列分析,并对所发现的点突变进行了基于限制性内切酶MspⅠ的PCR-RFLP分析。序列分析表明野猪与大白猪相比存在着4个SNPs;该基因在野猪的肝脏、肾脏、肺、大肠和肌肉中表达,在心脏、胃、脾、小肠和睾丸中不表达;对14头野猪的酶切多态性分析表明该突变位点是多态位点,并且3种基因型的分布符合Hardy-Weinberg定律。

Retinol-binding protein is a transporter protein, it specifically binds and carries retinol （ Vitamin A） from the liver to target tissues, helps VA to come true the transport and metabolism in cell, and is important in educing physiologic function of VA. It is not only a candidate gene of reproductive performance, but also a candidate gene of research on VA metabolic block and obesity. In order to further reveal its genetic variability in population, and to find new genetic markers, RBP4 gene of wild boar （Sus scrofa ussuricus） was cloned and analyzed. The polymorphic analysis on the newly found variant was done with PCR-RFLP using the endonuclease Msp I. Sequence analysis showed that there were four SNPs between wild bear and yorkshire. It expressed in liver, kidney, lung, large intestine and muscle, but not in heart, stomach, spleen, small intestine and testiculus. Polymorphic analysis made clear that the variant was polymorphic site, and that the distribution of three genotypes conformed to the Hardy-Weinberg law.