Miao Xue-Xia, Xub Shi-Jie, Li Ming-Hui, Li Mu-Wang, Huang Jian-Hua, Dai Fang-Yin, Marino Susan W, Mills David R, Zeng Peiyu, Mita Kazuei, Jia Shi-Hai, Zhang Yong, Liu Wen-Bin, Xiang Hui, Guo Qiu-Hong, Xu An-Ying, Kong Xiang-Yin, Lin Hong-Xuan, Shi Yao-Zhou, Lu Gang, Zhang Xianglin, Huang Wei, Yasukochi Yuji, Sugasaki Toshiyuki, Shimada Toru, Nagaraju Javaregowda, Xiang Zhong-Huai, Wang Sheng-Yue, Goldsmith Marian R, Lu Cheng, Zhao Guo-Ping, Huang Yong-Ping
Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
Proc Natl Acad Sci U S A. 2005 Nov 8;102(45):16303-8. doi: 10.1073/pnas.0507794102. Epub 2005 Nov 1.
We established a genetic linkage map employing 518 simple sequence repeat (SSR, or microsatellite) markers for Bombyx mori (silkworm), the economically and culturally important lepidopteran insect, as part of an international genomics program. A survey of six representative silkworm strains using 2,500 (CA)n- and (CT)n-based SSR markers revealed 17-24% polymorphism, indicating a high degree of homozygosity resulting from a long history of inbreeding. Twenty-nine SSR linkage groups were established in well characterized Dazao and C108 strains based on genotyping of 189 backcross progeny derived from an F(1) male mated with a C108 female. The clustering was further focused to 28 groups by genotyping 22 backcross progeny derived from an F(1) female mated with a C108 male. This set of SSR linkage groups was further assigned to the 28 chromosomes (established linkage groups) of silkworm aided by visible mutations and cleaved amplified polymorphic sequence markers developed from previously mapped genes, cDNA sequences, and cloned random amplified polymorphic DNAs. By integrating a visible mutation p (plain, larval marking) and 29 well conserved genes of insects onto this SSR-based linkage map, a second generation consensus silkworm genetic map with a range of 7-40 markers per linkage group and a total map length of approximately 3431.9 cM was constructed and its high efficiency for genotyping and potential application for synteny studies of Lepidoptera and other insects was demonstrated.
作为一项国际基因组计划的一部分,我们利用518个家蚕(经济和文化上重要的鳞翅目昆虫)的简单序列重复(SSR,或微卫星)标记构建了一个遗传连锁图谱。使用基于2500个(CA)n和(CT)n的SSR标记对六个代表性家蚕品系进行的调查显示,多态性为17%-24%,这表明由于长期近亲繁殖导致了高度的纯合性。基于189个回交后代(由F(1)雄性与C108雌性交配产生)的基因分型,在特征明确的大造和C108品系中建立了29个SSR连锁群。通过对22个回交后代(由F(1)雌性与C108雄性交配产生)进行基因分型,聚类进一步集中到28个组。借助可见突变以及从先前定位的基因、cDNA序列和克隆的随机扩增多态性DNA开发的酶切扩增多态性序列标记,这组SSR连锁群被进一步分配到家蚕的28条染色体(已建立的连锁群)上。通过将可见突变p(普通,幼虫斑纹)和29个昆虫的高度保守基因整合到这个基于SSR的连锁图谱上,构建了第二代家蚕共识遗传图谱,每个连锁群有7-40个标记,总图长约3431.9 cM,并证明了其在基因分型方面的高效性以及在鳞翅目和其他昆虫的同线性研究中的潜在应用。