民猪Haln基因部分序列片段比对分析

摘要

To determine halothane gene nucleotide sequence, genetype frequency and gene frequency in minpig, using polymerase chain reaction (PCR) , the halothane gene (Hal") of Northeast minpig include ing complete exon 17 and partial introns 16 and 17 was amplified. The amplification products were under taken enzyme cut polymorphism analysis using Hha I incision enzyme, minpig HalNHalN(NN) type was found to be 70.83% , HalNHaln(Nn) type was 25% , Hal" HalN(nn) type was 4.17% ; N gene frequen cy was 83.33% and n gene frequency was 16.67% . The 612 bp halothane gene fragment was se-quenced, and homology analysis showed that minpig halothane gene sequence had 99% homology with halothane gene sequence in GenBank and the presumed amino acids sequence homology was 95% . By comparing minpig halothane gene nucleotide sequence to 11 known domestic and overseas varieties of pig, 15 mutation sites were found. Minpig and Tibetan pig were found to have the neariest genetic distance and constitute a branch by doing phylogenetic analysis between 12 varieties (strains) pig halothane gene. The other varieties' pig halothane gene had closer genetic variation, overlapping phenomena. This re search identified halothane gene genetic variation among different varieties of pigs, providing theoretical foundation for varietal improvement and for local variety pigs.%为确定民猪群体中Haln基因的核苷酸序列、基因型频率及基因频率,采用PCR方法体外扩增了民猪Haln基因完整的外显子17和内含子16,17的部分序列.采用HhaⅠ内切酶对扩增产物进行了酶切多态性分析,发现民猪HalN HalN( NN)型占70.83％,HalNHaln(Nn)型占25％,HalnHaln(nn)型占4.17％;N基因频率为83.33％,n基因频率为16.67％.对所获得的序列长度为612 bp的Haln基因片段进行序列测定同源性分析.结果表明:民猪与GenBank中已收录的Haln基因的同源性为99％,所推测的氨基酸序列同源性为95％.对民猪Haln基因核苷酸序列与已知国内外11个品种猪Haln基因核苷酸序列进行比对,共检测到15个碱基突变, 在对12个品种(系)猪Haln基因进行系统发生分析时发现,民猪与藏猪的Haln基因的遗传变异最小,构成1个支系,其他各品种间Haln基因的遗传变异较接近,彼此间有交叉现象.