摘要
[目的]建立检测马MxA基因第12外显子多态性的快速、准确方法。[方法]采用错配聚合酶链反应(mismatch PCR)对马MxA基因第12外显子进行扩增,对PCR产物进行限制性片段长度多态性分析(RFLP),鉴定MxA基因cDNA第1 790位核苷酸点突变,并对PCR产物进行核苷酸序列分析。[结果]马MxA基因第12外显子区域存在AA、AB、BB3个基因型;位于cDNA序列第1 790位点的碱基发生变异(由G→C),引起了MxA蛋白编码区第562氨基酸由色氨酸变成半胱氨酸的变异;使用mismatchPCR-RFLP法所得PCR产物特异性序列,与RFLP分析结果相符。[结论]采用mismatch PCR-RFLP对马MxA基因12外显子的多态性进行检测操作简单,结果准确。
[ Objective ] The experiment aimed to establish a fast and accurate method to detect the polymorphism of No. 12 exon in MxA gene of horse. [Method] The mismatch PCR was used to amplify the No. 12 exon, then the PCR product was conducted RFLP. It was concluded from the identification that the nucleotide 1 790 site in cDNA of MxA gene was mutated and the PCR product was undergone nucleotide sequence analysis. [ Result ] There were 3 genotypes（ AA,AB, BB） in the region of No. 12 exon in MxA gene of horse. The bases of nucleotide 1 790 site in cDNA of MxA gene were mutated（ G was changed to C ） and this mutation caused the change of tryptophan to cysteine. The specific sequence of PCR-RFLP product deserved from mismatch PCR was same to that deserved from RFLP analysis. [ Conclusion ] The mismatch PCR-RFLP method was easy to detect No. 12 exon in MxA gene of horse and the result was accurate.
出处
《安徽农业科学》
CAS
北大核心
2009年第8期3463-3464,3467,共3页
Journal of Anhui Agricultural Sciences
基金
内蒙古自治区自然基金资助项目(200508010413)
