家蚕精氨酸激酶基因的克隆,基因结构及表达分析.pdfVIP

中国农业科学 2006,39(11):2354-2361 Scientia Agricultura Sinica 家蚕精氨酸激酶基因的克隆、基因结构与表达分析 王华兵，徐豫松 （浙江大学动物科学学院，杭州 310029 ） 摘要： 【目的】克隆家蚕精氨酸激酶基因，分析其基因结构与表达特性,为揭示无脊椎动物体内能量代谢调 节规律提供重要基础。【方法】通过分析家蚕EST、利用RACE法和基因组文库筛选法克隆了家蚕精氨酸激酶 （ BmAK, Bombyx mori arginine kinase）基因,并对其基因结构和表达特性进行了分析。【结果】克隆了BmAK 基因, cDNA 全长为1 268bp,编码355个氨基酸，具有精氨酸激酶典型的酶活性部位氨基酸序列，酶活性中心位点氨基酸和能 形成离子偶结构氨基酸；该基因由 2 个外显子和 1 个内含子组成；5′调控序列存在 BRCZ、E74A、FTZ 等多个潜在 转录因子结合位点，但没有TATA盒启动子序列；该基因的表达在不同组织和不同发育时期存在明显差异。【结论】 BmAK具有精氨酸激酶的典型特征，BmAK基因表达随发育时期不同而发生变化，基因的表达可能受蜕皮激素调控。 关键词：家蚕；精氨酸激酶；基因结构；表达；转录因子 cDNA Cloning, Genomic Structure and Expression of Arginine Kinase Gene from Bombyx mori (L.) WANG Hua-bing, XU Yu-song (College of Animal Sciences, Zhejiang University, Hangzhou 310029) Abstract: 【Objective 】In order to clarify regulation mechanisms of energy metabolism of invertebrates, the arginine kinase gene in the silkworm,Bombyx mori was cloned and analyzed. 【Method 】Based on the information of silkworm ESTs, the full-length cDNA of arginine kinase gene was cloned using the strategy of rapid amplification of cDNA ends, and then the genomic structure of BmAK was analyzed. Further, the expression of BmAK was examined. 【Results 】BmAK cDNA has 1268 bp and contains an open reading frame encoding a 355 amino acid protein. BmAK contains the active sequence, the active site and a pair of highly conserved amino acids that form an ion pair. The BmAK gene contains only 2 exons and 1 intron. The BmAK promoter contains no TATA box, but potential binding sites for the transcription factors in both forward and reverse orientation:BRCZ、E74A、FTZ,