›› 2011, Vol. 54 ›› Issue (11): 1223-1230.doi:

• 研究论文 • 上一篇    下一篇

家蚕第12连锁群EST-SSR标记的筛选和分析

 米智, 李傲祥, 阮成龙, 李冠楠, 杜文华, 隆耀航, 朱勇   

  • 收稿日期:2011-06-22 修回日期:2011-10-18 出版日期:2011-11-20 发布日期:2011-11-20
  • 通讯作者: 朱勇 E-mail:zhuy@swu.edu.cn
  • 作者简介:米智, 男, 1985年生, 山西大同人, 博士研究生, 研究方向为家蚕遗传育种, E-mail: mizhi775@126.com
  • 基金资助:

     重庆市蚕桑重大科技专项项目(CSTC, 2009AA1024)

Searching and analysis of EST-SSR markers from linkage group 12 of the silkworm, Bombyx mori

 MI  Zhi, LI  Ao-Xiang, RUAN  Cheng-Long, LI  Guan-Nan, DU  Wen-Hua, LONG  Yao-Hang, ZHU  Yong   

  • Received:2011-06-22 Revised:2011-10-18 Online:2011-11-20 Published:2011-11-20
  • Contact: ZHU Yong E-mail:zhuy@swu.edu.cn
  • About author:mizhi775@126.com

摘要: 为了探究家蚕Bombyx mori EST-SSR标记的多态性, 对检索获得的家蚕第12连锁群的4 465条EST序列进行了分析, 整理和拼接后得到581条非冗余EST序列, 总长度约为480 kb。其中, 有122条序列中共检测到154个EST-SSR, 占所研究的EST序列的2.73%, 平均每3.12 kb 含有一个EST-SSR。在所检测的EST-SSR中, 三核苷酸和四核苷酸重复是主导类型, 分别占总数的36.36%和28.57%,大部分表现为Perfect形式; 核苷酸重复平均长度约为16.2 bp, 最长为30 bp。进一步进行同源性分析, 发现有26条序列可以在NCBI中检索到同源序列, 在这些序列中一共含有40个SSR, 其中14个(35.0%)位于5′-UTR, 11个(27.5%)位于3′-UTR, 15个(37.5%)位于CDS区。根据筛选到的微卫星序列设计11对引物, 其中8对引物有扩增产物, 且条带清晰; 应用引物ES1204对8个家蚕品种进行PCR扩增都呈现多态性。结果说明通过家蚕EST数据库发掘SSR标记是一条可行的途径。

关键词: 家蚕, 连锁群, 表达序列标签, EST-SSR标记, 多态性

Abstract:  In order to explore the polymorphism of EST-SSR markers in silkworm, we retrieved and analyzed 4 465 expressed sequence tags (ESTs) on linkage group 12 of the silkworm, Bombyx mori. After splicing and jointing, we obtained 581 non-redundant ESTs with the total length about 480 kb. A total of 154 simple sequence repeats (EST-SSRs) were detected in 122 ESTs which accounted for 2.73% of all the investigated ESTs, meaning one EST-SSR per 3.12 kb on average. Trinucleotide repeats and tetranucleotide repeats were the main classes, which accounted for 36.36% and 28.57% of all detected EST-SSRs, respectively. The average length of nucleotide repeats was 16.2 bp, and the maximum was 30 bp. Homologous sequences of 26 EST-SSRs were retrieved in NCBI database, which contained 40 SSRs. Among them, 14 SSRs (35.0%) were located in the 5′-UTR, 11 SSRs (27.5%) in the 3′-UTR and 15 SSRs (37.5%) in the coding sequences (CDSs). Eleven primer pairs were designed for the screened EST-SSRs and 8 of them showed polymorphic bands. The primer ES1204 showed polymorphism among 8 silkworm strains. The results suggest that it is feasible to find SSR markers through searching of silkworm EST databases.

Key words: Silkworm (Bombyx mori), linkage group, expressed sequence tags (ESTs), EST-SSR markers, polymorphism