利用SSH技术鉴定黄瓜胚胎发育相关基因  

Identification of Embryo Development-Related Genes in Cucumber Using SSH

作　　者：盛慧[1] 秦智伟[1] 周秀艳[1] 武涛[1] 

机构地区：[1]东北农业大学园艺学院黄瓜分子育种实验室,哈尔滨150030

出　　处：《中国农业科学》2010年第11期2292-2299,共8页Scientia Agricultura Sinica

基　　金：国家"863"计划项目(2006AA100108-4);哈尔滨市科技创新人才资金项目(008RFXXN002)

摘　　要：【目的】寻找黄瓜胚胎发育早期和胚胎发育晚期优势表达基因,研究其胚胎发育的分子机制。【方法】利用抑制性消减杂交(suppression subtractive hybridization,SSH)技术构建胚胎发育早期和晚期差异表达cDNA文库。【结果】经反向Northernblot杂交检测,共得到97个阳性克隆。利用分子生物学软件对测序后的序列进行质量检测和聚类、拼接,共得到93个UniESTs,其中包括86个singletons和7个contigs。将上述EST序列在GenBank上进行BLAST比对,有83个EST片段找到了与之匹配的同源序列,有10个EST片段未找到同源序列,推测可能是新基因。【结论】其中很多EST与拟南芥、水稻或玉米蛋白质数据库中的热激蛋白具有很高的同源性。将以上序列应用GeneOntology进行功能分类,发现在胚胎发育早期细胞防御和代谢过程占主要位置;在胚胎发育晚期细胞防御和抗渗透胁迫功能是非常重要的。【Objective】 The aim of this experiment is to investigate the embryo development-related genes in cucumber,and study its molecular mechanism.【Method】 A subtracted cDNA library was constructed by using Suppression Subtractive Hybridzation （SSH）.【Result】 A total of 97 positive clones were screened using reverse Northern blot hybridization.After the removal of duplication and sequencing of low-quality sequences,93 UniESTs were identified,including 86 singletons and 7 contigs.The 93 unique ESTs were used to BLAST in the GenBank non-redundant database.There were 83 ESTs were found matching homologous sequences,10 ESTs homologous sequence were not found,presumably may be new genes.【Conclusion】 Many of these ESTs were highly homologous with the heat shock proteins of Arabidopsis,rice and corn.Application of the GeneOntology functional classification,cell defense and metabolic processes in early embryos accounted for the main location.Cell defense and anti-osmotic stress function is very important in late embryos.

关 键 词：黄瓜 SSH 胚胎发育 CDNA文库 

分 类 号：S642.2[农业科学—蔬菜学]