关中奶山羊PIS区域序列多态性分析

摘要

山羊无角性状是畜牧业生产中重要的经济性状,报道显示山羊无角间性综合征(polled intersex syndrome,PIS)控制着山羊的无角和间性性状.山羊PIS序列定位于1号染色体约11.7 kb的序列上,试验旨在研究这段序列对关中奶山羊(3只有角山羊、3只无角山羊)无角性状的影响.选用4对引物(PIS1 1、PIS1-2、PIS2、PIS3)扩增关中奶山羊的PIS序列,第5对引物(PIS whole)用于检测PIS序列的完全缺失,对测序结果进行拼接、多态性分析,并对序列进行基因注释.结果发现,试验成功扩增出山羊PIS的全长序列(12.814 kb),BLAST比对结果正确.PIS whole引物没有扩增出任何条带,说明关中奶山羊中不存在11.7 kb的完全缺失.经SeqMan软件分析拼接序列发现30个多态性位点,这些多态性位点可能与关中奶山羊有角/无角性状相关.对拼接序列进行基因注释,发现存在1个tRNA编码位点、2个微卫星、3个microRNA编码位点及L1-EN、RT-nLTR-like样保守结构域,并发现1个CpG岛、1对反向重复序列和2段串联重复序列.本试验结果对关中奶山羊无角性状的分子机理研究具有重要的参考价值.%Polled trait of goat is important economic trait in livestock production,goat polled inter sex syndrome (PIS) influenced polled phenotype and intersex phenotype of goats.PIS sequence locats in a approximately 11.7 kb nucleotide sequence in chromosome 1.This experiment was aimed to study the effect of PIS sequence on polled phenotype in Guanzhong dairy goats (3 horned goats,3 polled goats).The PIS whole sequence in Guanzhong dairy goats were amplified using four pair of primers (PIS1-1,PIS1-2,PIS2 and PIS3),the whole absence of PIS sequence was detected using the fifth primer (PIS whole).The sequencing result was assembled,analyzed and annotated.The result showed that the PIS sequence full length was 12.814 kb,and the BLAST alignment result was correct.PIS whole primer didn't amplify any srtipes,which showed that there was no a total deletion of 11.7 kb in Guanzhong dairy goats.SeqMan software analysis result showed that there was a total of 30 polymorphic sites,wihch were likely to connect with polled/horned phenotype in Guanzhong dairy goats.The annotation result of concatenation sequence showed that there were one tRNA encoding locus,two microsatellites,three microRNA encoding loci,L1-EN conserved domain,RT-nLTR-like conserved domain,also find a CpG island,one pair of inverted repeats,and two tandem repeats.This experiment result had a significant reference value for studying the molecular mechanism of polled phenotype in Guanzhong dairy goats.