基于HRM分析技术的家蚕眠性主基因M的精细定位  

Fine Mapping of Major Moltinism Gene M in Bombyx mori Based on HRM Analysis Technique

作　　者：刘晓晓[1] 张洁葵[1] 张业顺[2] 杨思思[1] 夏定国[2] 余俊杰[1] 刘丽丽[1] 吴阳春[2] 张国政[1,2] 

机构地区：[1]江苏科技大学,江苏镇江212003 [2]中国农业科学院蚕业研究所,农业部蚕桑遗传改良重点实验室,江苏镇江212018

出　　处：《蚕业科学》2013年第4期656-662,共7页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31272499);江苏省研究生科技创新计划项目(No.CXZZ12_0727)

摘　　要：就眠蜕皮是家蚕幼虫的一种重要生理现象,家蚕眠性性状主要受第6连锁群眠性主基因M控制。用四眠蚕品种大造(p50)与三眠蚕品种C3白构建BC1M群体p50×(C3白×p50),选择其中的598头三眠蚕个体作为定位群体,应用高分辨率熔解曲线(HRM)分析技术进行家蚕基因组SSR标记基因型分析,对M基因进行精细定位。通过HRM筛选出8个与M基因连锁的多态性SSR标记,据此绘制出遗传总距离为2.4 cM的分子连锁图,标记S2853-2527和S2853-2843位于M基因两侧,遗传距离分别为0.2 cM和0.7 cM,M基因位于家蚕基因组nscaf2853:2527929～2843864之间,其间有4个预测基因都属于序列特异的DNA结合转录因子,推测眠性主基因的作用方式可能与DNA序列特异性转录调控有关。研究结果也验证了HRM技术应用于家蚕SSR标记基因型分析的可行性和可靠性。Molting and metamorphosis of silkworm（Bombyx mori） larvae is an important physiological phenomenon.Moltinism trait of silkworm is mainly controlled by major moltinism gene M which is located on the 6th linkage group.In this study,we used a tetramolter silkworm race p50 and a trimolter silkworm race C3-white as parental strains to prepare BC1M population p50×（C3-white×p50）.598 trimolter individuals of BC1M were selected as gene mapping population,and genotyping analysis of genomic SSR molecular markers was employed for fine mapping of M gene by using high-resolution melting（HRM） analysis technique.By HRM analysis technique,we obtained eight silkworm polymorphic SSR markers linked with M gene,and constructed molecular linkage map of M gene with total genetic distance of 2.4 cM.SSR marker S2853-2527 and S2853-2843 were located at the two sides of M gene,having a genetic distance of 0.2 cM and 0.7 cM with M gene respectively.M gene locates between nscaf2853：2527929 and nscaf2853：2843864 of silkworm genome,in which there are 4 predicted genes.All these genes encode sequence-specific DNA binding transcription factors,suggesting that M gene has similar functioning mechanism with them.Our results demonstrated the feasibility and reliability of using HRM analysis technique to conduct genotyping analysis of silkworm SSR markers.

关 键 词：家蚕 眠性主基因 高分辨率熔解曲线分析 SSR标记 基因定位 

分 类 号：S881.26[农业科学—特种经济动物饲养]