猪锌转运蛋白基因的克隆及其在消化道组织中的分布

摘要

slc30A family (vertebrate cation diffusion facilitator family proteins) transport zinc from the cytoplasm to the lumen, thus decreasing the content of zinc in cytosol. Cloning primers were designed according to the cloning principle of homologous sequence in this paper, the slc30A l~slc30A 9 genes were cloned by RT-PCR, then compared cloning sequences of slc30A1~slc30A9 and protein amino acid coding by these gene sequences with other species for homology analysis, and we also studied the slc30Al~slc30A9 distribution in the pig tissue. The main results showed that: The sequence length of gene slc30Al~slc30A9 was about 1 044-2 301 bp all with a complete open reading frame (ORF), which medicated to encode 348~767 amino acid residues, respectively(GenBank accession No. FJ374262.1, EU825192.2, FJ358706.1, EU835903.1, FJ230771.1, FJ237622.1, FJ237623.1, FJ588029 and FJ164069.1, respectively), but slc30A8 was the only mutant. The homology analysis results of slc30A and ZnT (zinc transporter) protein amino acid nearly were the same, higher with human (Homo taurus) and cow (Bos taurus), but lower with Rat (Rattus nurvegicus) and Mouse(Mus musculus), and difference were present in the different isomers and its coding proteins. The results of tissue distribution suggested that slc30A8 was only found in the ileum, slc30A9 was found in the esophagus and all tract, while slc30A 1 and slc30A 6 were found in other tissue except in the ileum. The other isomers were not found in the ileum, but there were different degree distribution in other tissue. These results suggested that the difference of the slc30Al~ slc30A9 genes distribution in all tissues maybe related to its special function in pig tissue.%slc30A编码的锌转运蛋白可将胞质锌排出细胞外或转入细胞器,从而降低胞浆锌的含量.本研究运用电子克隆设计引物,对猪(Sus scrofa)消化道组织中slc30A l～slc30A9进行克隆,将克隆的slc30A1～slc30A9序列及其编码蛋白的氨基酸序列与其他物种进行同源性比较,并对slc30A1～slc30A9在猪消化道组织中的分布进行研究.克隆结果表明,猪slc 30A1～slc 30A9均包含一个完整的开放阅读框架,长度为1 044～2 301 bp,编码由348～767个氨基酸残基组成的ZnT(Zinc transporter)蛋白,GenBank登录号分别为:FJ374262.1、EU825192.2、FJ358706.1、EU835903.1、FJ230771.1、FJ237622.1、FJ237623.1、FJ588029和FJ164069.1,其中slc30A8为一个突变体；同源性分析比较发现,slc30A和ZnT蛋白氨基酸与其他种属的同源性分析结果基本一致,与牛(Bos taurus)和人(Homo sapieus)的同源性较高,与小鼠(Mus musculus)和大鼠(Rattusy norvegicus)的同源性较低,且不同异构体及其编码的蛋白也略有差异；组织分布结果表明,除slc30A8仅在回肠中有分布外,slc30A9分布最广,在食道、十二指肠、空肠、回肠、盲肠、结肠和直肠中均有分布,而slc30A1和slc30A6除了回肠,其余组织均有分布,其他几个异构体在回肠中也没有分布,而在食道、十二指肠、空肠、盲肠、结肠和直肠中有不同程度的分布.研究结果提示,slc30A在消化道中的分布具有组织特异性,可能与其在猪体内的功能密切相关.