来航鸡FOXL2基因的克隆与序列分析

Cloning and sequencing analysis of the FOXL2 Gene of Leghorn chicken

为了研究来航鸡FOXL2基因的结构及功能,根据GenBank上登录的红色原鸡FOXL2基因序列设计引物,对来航鸡FOXL2基因进行了克隆、测序,并对该序列进行生物信息学分析。结果表明:克隆得到的来航鸡FOXL2基因全长为1 130 bp(GenBank登录号为JF708868),包含1个918 bp的完整开放式阅读框,编码305个氨基酸,其CDS编码区的核苷酸序列与红色原鸡、人、牛、野猪、家鼠、欧洲兔、蟾蜍、斑马鱼及罗非鱼的FOXL2基因对应序列的同源性分别为99.8%、80.0%、80.4%、80.3%、80.5%、81.5%、77.7%、71.8%、75.5%,推导的氨基酸序列同源性分别为99.7%、64.7%、64.7%、65.4%、63.5%、63.8%、79.7%、78.3%、79.9%;FOXL2编码蛋白主要存在于细胞核中,存在1个保守结构域,不含信号肽、跨膜结构域和剪切位点,并存在2个蛋白质激酶C磷酸化位点、1个cAMP和cGMP依赖的蛋白激酶磷酸化位点、2个N-糖基化位点、3个酪蛋白激酶Ⅱ磷酸化位点和4个N-豆蔻酰化位点,预测最佳抗原表位候选区域为45～49位(SQKPP)、147～152位(RPPPTH)和169～173位(SPPKY)。

To study the structure and function of FOXL2 gene of Leghorn chicken, the primers were designed based on the sequence of the FOXL2 gene sequences of Gallus gallus in GenBank. The FOXL2 gene was cloned by PCR and the sequence was analyzed using the bioinformat- ic tools. The results showed that the FOXL2 gene of Leghorn chicken was 1 130 bp （GenBank accession： JF708868） in length with a 918 bp open reading frame and encoded 305 amino acids. The nucleotide sequence of CDS shared 99.8% ,80.0% ,80.4% ,80.3% ,80.5% ,81.5%, 77.7% ,71.8% and 75.5% homology with the FOXL2 mRNA of Callus gallus, Homo sapiens, Bos Taurus, Sus scrofa, Mus musculus, Orycto- lagus cuniculus, Xenopus laevis, Danio rerio and Oreochromis niloticus respectively, and the homology of deduced amino acids were 99.7%, 64.7% ,64.7% ,65.4% ,63.5% ,63.8% ,79.7% ,78.3% and 79.9% respectively. It was predicted that the encoded protein of FOXL2 gene mainly existed in the nucleus, which contained one conserved domain, but it had no signal peptide, transmembrane region and splice sites. It contained two protein kinase C phosphorylation sites, one cAMP and cGMP dependent protein kinase phosphorylation site, two N - glycosylation sites, three casein kinase 11 phosphorylation sites and 4 N - myristoylation sites. It was predicted that the best candidate regions for the antigen epitope were located at the regions of 45 - 49 （SQKPP） , 147 - 152（RPPPTH） and 169 -173 （SPPKY）.