摘要
以番茄抗病品系（CLN2037E）为研究材料,应用SMART技术构建晚疫病诱导的番茄叶片cDNA表达文库,并进行生物信息学分析。成功构建的cDNA文库的克隆数为2.3×109,重组率为96%,通过PCR检测,插入片段范围在0.1-2.0 kb之间。从文库中随机挑取110个克隆进行测序,最终得到107条差异表达ESTs序列。GenBank提交的序列号为GT742146-GT742150;GT865998-GT866099。利用BLAST进行同源性分析：75条ESTs与其他物种同源性较高,视为已知基因,功能涉及能量代谢的占42.7%、细胞内生命活动与信号传导的占10.7%、基因转录和翻译的占9.3%以及与抗性相关的占25.3%;另有32条ESTs无同源序列或同源性较低,预测是一些未知功能基因的占30%。本项研究应用SMART技术成功构建了高质量、信息丰富的晚疫病诱导的番茄叶片cDNA文库,为进一步筛选抗病相关候选基因奠定了基础。
A cDNA library was constructed to investigate late blight disease-resistant mechanism and initially obtained some disease resistant related Expressed Sequenced Tags（ESTs） of tomato（Lycopersicon esculentum Mill.） leaves infected by late blight disease（Phytophthora infestans） using SMART（Switching mechanism at 5′ end of RNA transcript） technique.The library had a high titer of 2.3×109 pfu/ml with a recombination rate of 96%.The inserted cDNA fragments ranged from 0.1～2.0 kb by PCR detection.After differential screening on the cDNA library,107 ESTs were identified and their GenBank accession numbers were GT742146-GT742150 and GT865998-GT866099.BLAST analysis showed that：75 ESTs presented significant similarity to the genes identified from other species.It was found that 42.7% of the ESTs involved in energy metabolism,10.7% in cellular life activities and signal conduction,9.3% in gene transcription and translation and 25.3% of the ESTs was involved in resistance.32 ESTs with low similarity or no match to the known genes,and they may contain some unidentified functions.The cDNA library enjoyed good quality and rich information,and the information generated in this study had established a basis for screening candidate genes involved in tomato disease-resistant.
出处
《中国农业大学学报》
CAS
CSCD
北大核心
2010年第5期65-70,共6页
Journal of China Agricultural University
基金
云南省科技攻关计划(2006NG17)
关键词
番茄
晚疫病
CDNA文库
EST序列分析
tomato
late blight disease（Phytophthora infestans）
cDNA library
Expressed Sequenced Tags analysis
