基于比较基因组学方法,选择大白猪和梅山猪作为试验材料,根据人、小鼠和猪的BMP15基因设计并合成4对引物,进行基因组DNA的PCR扩增、克隆、测序,用BLAST软件进行DNA序列排列,获得包含猪BMP15基因外显子1(exon1)和外显子2(exon2)的全部编码区序列。用Pairwise BLAST软件,将大白猪、梅山猪BMP15基因编码区序列进行比较,在外显子2区域发现了一个SNP位点,位于编码区第390个核苷酸处,大白猪为T,梅山猪为A,且该位点导致了限制性内切酶SpeⅠ酶切位点发生了改变。建立了猪BMP15基因基于内切酶SpeⅠ的PCR—RFLP多态性检测技术,发现猪BMP15基因有3种基因型(BMP15^AA、BMP15^AB、BMP15^BB)。
This study was established on comparing genomics using Duroc, Large White and Meishan pig as materials. According to the BMP15 sequeces of human, rat and pig, four pairs primers were designed and synthesized by comparing genomics. The genome DNA was amplified using the primers. Then, the product was cloned and sequenced. After that, the DNA sequences were sequenced using BLAST soft . The complete coding sequences including exon 1 and exon 2 of pig BMP15 were obtained. Comparing the coding sequences of Large White and Meishan pig BMP15 by Pairwise BLAST, a new SNP was found in the area of exon 2. The SNP was at the point of 390^th nucleotide of the coding sequences. At this point, the nucleotide of Large White pig was T while Meishan pig of that was A. The SNP resulted in the change of the digesting site of restriction endonuclease Spe Ⅰ . BMP15 had three genotypes : BMP15^AA, BMP15^AB and BMP15^BB.