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单链构象多态性(Single strand conformation polymorphism,SSCP)技术是一种简便、灵敏的多态性检测方法,可以检测出在非变性聚丙烯酰胺凝胶电泳中因构象差异而导致的单链DNA片段迁移率的不同。本研究根据棉花基因芯片筛选的纤维发育中差异表达基因设计了162对引物,利用SSCP技术在4个陆地棉品种、4个海岛棉品种中进行多态性检测。结果表明,在162对引物中,146对引物经PCR扩增后在1.5%的琼脂糖凝胶电泳中检测出现清晰、明亮的带。经过SSCP分析,54对引物在陆地棉之间产生多态性,共出现116个多态性位点;45对引物在海岛棉之间产生多态性,共出现111个多态性位点;79对引物在陆地棉和海岛棉之间产生多态性,共出现260个多态性位点;36对引物在陆地棉之间、海岛棉之间同时出现多态性。进一步聚类分析后表明,海岛棉和陆地棉分别聚在了一起。
Single strand conformation polymorphism (SSCP) is a simple and sensitive method to detect single-stranded DNA (DNA) due to conformational differences in non-denaturing polyacrylamide gel electrophoresis Different fragment mobility. In this study, 162 pairs of primers were designed based on the differentially expressed genes in the fiber development screened by cotton gene chips. The SSCP technique was used to detect the polymorphism in 4 upland cotton varieties and 4 sea island cotton cultivars. The results showed that out of 162 pairs of primers, 146 pairs of primers were detected by 1.5% agarose gel electrophoresis with clear and bright band after PCR amplification. After SSCP analysis, 54 pairs of primers produced polymorphism among G. hirsutum and a total of 116 polymorphic loci were observed. Forty-five pairs of primers produced polymorphism between G. barbadense and 111 polymorphic loci. A total of 260 polymorphic loci were found in 79 pairs of primers between G. hirsutum and G. barbadense. Thirty-six pairs of primers were polymorphic between L. gossypii and G. gossypii. Further clustering analysis showed that sea-island cotton and Upland cotton were clustered together.