摘要
柠条是包括内蒙古在内干旱荒漠草原上的优势建群植物,具有极强的耐旱(寒)力,为了研究其抗逆机理,挖掘和利用其抗旱基因,用Trizol法从经过干旱处理的柠条当年新生幼嫩枝叶中提取总RNA,分离纯化mRNA,采用SMART技术反转录合成双链cDNA,DSN技术进行均一化,构建了柠条全长cDNA文库。经检验,文库的滴度为1.5×106 cfu/mL,重组率为93.75%,90%以上插入片段的长度为1.0～2.0 kb。从文库中随机挑取3200个克隆进行5'测序,得到3020个有效EST,合成2429个unigenes,其中,singlets 2229条,占总有效序列的91.76%,经与数据库比对,发现了脱水素、抗旱相关转录因子等多个抗旱相关基因。
In order to study the drought tollerence genes of korshinsk peashrub,a normalized full-length cDNA library was constructed using Creator SMART technology combined with duplex-specific nuclease（DSN） from water stressed leaves.The titer of original library was 1.5×106 cfu/mL,the recombinant efficiency was 93.75% and the average of insert size was over 1 kb,these results showed the cDNA library was of high quality.Besides,3 200 clones from it were selected randomly and sequenced.2 429 unigenes were obtained,among them 2 229 was singlets,means this library was normalized well.Some genes which response to drought were found after BLAST.
出处
《华北农学报》
CSCD
北大核心
2012年第3期67-71,共5页
Acta Agriculturae Boreali-Sinica
基金
内蒙古农牧业科学院创新基金(2007CXJJN03)
