【作者】 塔娜；

【导师】 芒来；

【作者基本信息】 内蒙古农业大学 ， 动物遗传育种与繁殖学， 2008， 硕士

【摘要】 本研究以不同类型西南矮马为研究对象,参考着GenBank中已发表物种的矮小性状基因SHOX的序列,利用PCR技术,首次克隆了7个类型西南马矮小性状基因SHOX的部分序列。其中,德保马的序列提交到了GenBank中,登录号为:EU556739。此外,对7个类型西南马的矮小性状基因序列及核苷酸组成进行了初步研究,同时采用PCR-SSCP技术和PCR产物直接测序等方法,采用纯种设特兰矮马(ST,3只)作为对照组,对各类型西南马SHOX基因序列进行了多态性分析。研究结果表明:1、克隆所得的序列长度为520bp,包含着CDS的部分序列。2、通过GenBank中的Blast软件对此序列进行同源性比较以后发现,与人的矮小性状基因SHOX有95%的相似性。并通过DNAman等软件比较发现此片段主要是马矮小性状基因SHOX的第2外显子部分。3、将序列提交到了GenBank,登陆号为EU556739。4、西南马SHOX基因序列核苷酸组成富含GC,且马与其它物种的SHOX基因密码子在第2外显子上核苷酸均偏向使用G和C。5、PCR-SSCP分析结果表明,在克隆所得序列上选择的两个位点中P2位点出现了多态性,经测序得知在第268处有一个G→A的突变,在431处有一个T→G的突变,由此导致等位基因B变为等位基因A。该位点除云南文山马的优势基因型和优势等位基因为BB型和B外,其余类群马均为AB型,且X2检验结果表明:云南丘北马、云南文山马、云南大理马、建昌马和设特兰马在P2位点上处于Hardy-Weinberg平衡状态。6、不同类型马SHOX基因P2位点的纯合度(Ho)、杂合度(He)、有效等位基因数(Ne)及多态信息含量(PIC)分析表明:云南文山马的多态信息含量最高,并且处于高度多态(PIC>0.5),其它类型马(除设特兰马外)均为中度多态。更多还原

【Abstract】 Referencing by the SHOX gene sequence in GenBank, Seven types of Xinan pony SHOX gene sequence containing partial CDS sequence were firstly cloned and sequenced using the PCR technique.The sequence of DB horse was submitted to GenBank.Its accession is EU556739.The compositions of nucleotide of SHOX gene was analysed.The polymorphisms of SHOX gene of different horse groups were analyzed by PCR-SSCP and directly sequencing of PCR products. The results were as follows:1、The sequence of cloned pony SHOX gene was 520bp in length, included partial CDS sequence.2、The homology analysis of this sequence by BLAST software in GenBank showed that this sequence has 95% identity with homo sapiens SHOX gene sequence, and analysis by the DNAman indicated that this region was mainly Exon2.3、Submitted this sequence to the GenBank, and its accession number is EU556739.4、The analysis of nucleotide composition showed that G/C content was abundant in horse SHOX gene,and in exon 2 of different species,the bases of biased were all G and C.5、The analysis of PCR-SSCP showed that P2 site existed variations. The transversion of G→A in 268 and T→G in 431 led allelomorphic gene B→A. The dominant allele and genetype of WS were BB type and B, and the dominant allele and genetype of other horse types were AB. P2 site was in Hardy-Weinberg equilibrium in QB horse, WS horse ,JC horse and ST horse (P>0.05).6、Analysis result of the genetic polymorphism parameters of different horse groups at P2 site showed: Polymorphism of WS was very abundant at this site.更多还原