玉米sbe1和bt2基因SNPs位点的筛选

摘要

[目的]本研究针对玉米淀粉合成相关基因sbe1和bt2获得SNPs位点并设计引物,进行HRM-PCR分型,根据结果找到相关基因对玉米淀粉合成方面的调控机理,从而为玉米育种的遗传改良和分子设计育种提供理论参考.[方法]本文提取普通玉米H21和紫糯96-619的基因组DNA后进行PCR扩增,测序并拼接出玉米淀粉合成相关基因bt2,sbe1的全长序列.[结果]进行亲本间序列比对,找到了20个SNPs位点,通过高分辨率熔解曲线(HRM),成功对171个高世代回交群体的SNPs位点分型.[结论]运用试验分析,总结了利用直接测序法和HRM进行SNPs位点的筛选和验证的优缺点,以期为大规模SNPs筛选技术提供参考.%[Objective] Primers were designed based on the differerence of sequence alignment of sbe1 and bt2,according to the results of HRM-PCR,the regulation and control mechanism of candidate genes related to maize grain starch synthesis were found to provide a theoretical reference for genetic improvement and molecular design breeding of maize.[Method]The entire sequence of bt2,sbe1 in H21 and Zinuo96-619 by sequencing of PCR products and splicing were obtained.[Result] Sequence alignment results of genes between two parents showed that there were 20 SNPs.By means of high resolution melting (HRM) technology,171 individuals in BC 5 F2∶3 population were genotyped.[Conclusion] The advantages and disadvantages of sequencing method and HRM technology were summarized in order to provide a reference for large-scale SNPs screening technology in this paper.