昆虫学报2011，Vol.54Issue(1)：9-19,11.
家蚕生物钟基因Bmcryl与Bmcry2的克隆及生物信息学分析
Molecular cloning and bioinformatic analysis of biological clock genes Bmcry1 and Bmcry2 in Bombyx mori
摘要
Abstract
Cryptochrome gene (Cry) is one of the major biological clock genes which were widely distributed in bacteria and eukaryotes. Cry genes of insect species are clearly divided into two types, Cry1 and Cry2. Only Cry1 is expressed in Drosophila, while only Cry2 was expressed in bees and other hymenopteran insects. In order to explore the molecular mechanism of circadian clock in lepidopteran model insect Bombyx mori and the evolution of CRY proteins in insect species, we cloned the eDNA sequences of Bmcryl (2 166 bp, GenBank accession no. HM747059) and Bmcry2 (2 389 bp, GenBank accession no.HM747060), and obtained their gene sequences (GenBank accession no. HM747057 and HM747058,respectively) by sequence alignment and assembly. Bmcry1 have 12 exons and 11 introns, while Bmcry2 have 9 exons and 8 introns. Chromosome mapping showed that Bmcry1 and Bmcry2 were located on chromosome 17 and chromosome 15, respectively. We predicted the three-dimensional structure of Bmery1 and Bmcry2 by homology modeling. The results showed that the FAD entrances are large and deep,consistent with the fact that CRY proteins do not bind with pyrimidine dimers, and the surfaces of Bmcryland Bmcry2 are more negatively charged, only FAD entrances have accumulated positive charge. Moreover,we researched the molecular evolution of Bmcry1 and Bmcry2 by multiple sequence alignment, protein motif analysis, functional domain analysis and cluster analysis. The results showed that Bmcry1 and Bmcry2 belonged to insect CRY1 and CRY2, respectively, and were closely to the corresponding proteins in other lepidopteran insects like Antheraea pernyi. Similar to the CRY proteins in other insects, Bmcry1 and Bmcry2 have DNA photolysis enzyme domain and FAD binding domain. However, the two domains have different conservative sites between CRY1 and CRY2 in insect species, and their protein motifs are also different. Our experiment provided a basis for further investigating the mechanism of molecular evolution and function of CRY1 and CRY2 in B. mori.关键词
家蚕/生物钟/隐花色素基因/生物信息学分析/分子进化/同源建模Key words
Bombyx mori/ biological clock/ cryptochrome gene/ bioinformatic analysis/ molecular evolution/ homology modeling分类
生物科学引用本文复制引用
王文栋,梁辉,朱晓苏,陶卉,徐丽,司马杨虎,徐世清..家蚕生物钟基因Bmcryl与Bmcry2的克隆及生物信息学分析[J].昆虫学报,2011,54(1):9-19,11.基金项目
国家自然科学基金项目(30771632) （30771632）
国家重点基础研究发展规划("973"计划)项目(2005CB121005) （"973"计划）
国家科技支撑计划重点项目(2007BAD72801) （2007BAD72801）
苏州大学重大应用研究培育项目(Q3034850) （Q3034850）
