陆地棉GhLipase基因的克隆、特征分析及定位  

Molecular Cloning,Characterization and Mapping of GhLipase Gene in Gossypium hirsutum

作　　者：贺亚军[1] 郭旺珍[1] 张天真[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室棉花研究所,南京210095

出　　处：《农业生物技术学报》2009年第1期84-89,共6页Journal of Agricultural Biotechnology

基　　金：国家重点基础研究发展规划(973)项目(No.2002CB111301);长江学者和创新团队发展计划(No.IRT0432)资助

摘　　要：利用已构建陆地棉7235纤维cDNA文库测序,结合5′RACE技术从陆地棉(Gossypium hirsutum L)7235纤维中获得了1个1 286 bp的cDNA。序列分析结果表明,该cDNA包含一个编码368个氨基酸的开放读码框,5′非翻译区和一个带有Poly(A)的3′非翻译区区域。与已报道的其它植物的脂肪酶基因具有较高的氨基酸序列同源性,命名为GhLipase,提交到GenBank,登录号为EU273298。RT-PCR分析表明,GhLipase在棉花的胚珠及纤维细胞中优势表达。Southern blot分析结果表明,GhLipase在陆地棉基因组可能存在两个拷贝。将该基因定位在四倍体棉花遗传图谱的A13染色体上。A cDNA library was constructed using fibers of Gossypium hirsutum 7235 line. The library was randomly sequenced and an expressed sequence tag （EST） with the function of lipase was obtained. The full-length cDNA encoding lipase in 7235 line was cloned by the rapid amplification of eDNA ends （RACE） technique and named GhLipase. Sequence analysis showed that the full-length cDNA was 1286 bp which contained a major open reading frame of 368 amino acids, 5＇ un-translated regions, 3＇ un-trans-lated regions, and a Poly （A） tail. The sequence was submitted to GenBank, and accession No.EU273298. It had high homology to reported lipase genes from other plants. RT-PCR analysis confirmed that the GhLipase gene was up-regulated in ovules and fibers compared with roots, stems, and leaves of 7235. Southern blot analysis showed that there were two copies of the GhLipase gene in the genome of G. hirsutum. GhLipase gene was mapped on chromosome A13 in [（TM-1×Hai7124） TM-1] population.

关 键 词：棉花 脂肪酶基因 克隆 表达分析 基因定位 

分 类 号：S188[农业科学—农业基础科学]