南方农业学报2012，Vol.43Issue(6)：733-737,5.DOI:10.3969/j:issn.2095-1191.2012.06.733
干旱胁迫下甘蔗苗期根系全长cDNA文库构建及EST序列分析
Construction of full-length cDNA library and its EST sequence analysis in sugarcane root at seedling stage under drought stress
摘要
Abstract
[Objective]The present experiment was conducted to construct cDNA library of sugarcane root length and sequencing to provide referrence on isolation and clone of expressed drought genes. [ Method ] Under drought stress conditions, cDNA library of ROC22 sugarcane root was constructed using SMART technology and its sequence was analyzed. [Result]The identification results of library construction quality showed that the library storage capacity was l.0×1.06 PFU/ mL and recombination rate was about 95%, the inserts length was about 500 to 2000 bp and the average length was approximately 1102.1 bp. After unidirectional sequencing of 526 positive clones, a total of 523 ESTs sequences was obtained. Among them, a total of 468 Uni-ESTs was obtained by cluster analysis and assembling after removing vector, adapter sequence and the sequence with length less than 100 bp, in which included 29 congtigs and 439 singlets, accounting for 6.5% and 93.5% of the total, respectively. With the NCBI non-redundant protein database BLASTX comparasion, 444 Uni-ESTs had homologous sequences in the GenBank, which accounted for 94.87%; 24 unknown protein functional genes was also obtained, which accounted for 5.13%. [Conclusion]The present research contructed cDNA library of sugarcane root length under drought stress,and obtained 523 ESTs sequences,444 Uni-ESTs and a batch of unknown protein functional genes.关键词
cDNA文库/甘蔗/根系/表达序列标签/测序Key words
cDNA library/ root/ sugarcane/ expressed sequence tags/ sequencing分类
农业科技引用本文复制引用
罗海斌,曹辉庆,魏源文,邓智年,潘有强,郭元元,何海波,李杨瑞..干旱胁迫下甘蔗苗期根系全长cDNA文库构建及EST序列分析[J].南方农业学报,2012,43(6):733-737,5.基金项目
广西自然科学基金创新研究团队项目(2011GXNSFF018002) （2011GXNSFF018002）
广西科学研究与技术开发计划项目(桂科攻10100005-9) （桂科攻10100005-9）
广西科学基金项目(桂科基0778006-6,桂科基0832026,2010GXNSFA013102) （桂科基0778006-6,桂科基0832026,2010GXNSFA013102）
广西科学基金项目应用基础研究专项项目(桂科基0991021) （桂科基0991021）
广西农业科学院基本科研业务专项项目[200803Z(基),200902Z(基)] （基）
广西农业科学院科技发展基金项目(2007023) （2007023）
广西重点实验室建设项目(12-071-09) （12-071-09）
