枳根cDNA全长文库的构建与分析

Construction and Analysis of a Root Full-length c DNA Library of Poncirus trifoliate

为了解柑橘优良砧木枳(Poncirus trifoliata)根部的基因表达信息,利用SMART技术构建了枳的根组织全长c DNA文库。检测结果显示所建枳根原始文库的容量为1.08×106 cfu·m L-1,扩增后文库容量为1.30×109 cfu·m L-1,重组率为95%。通过文库随机测序获得182个长度大于100 bp的ESTs序列(Gen Bank登录号为JK316116~JK316297),序列拼接后得到96个Unigenes,包括12个Contigs和84个singletons。其中,68个Unigenes可与Gen Bank中登录的基因相匹配,56个与已知的柑橘基因相匹配,36个为全长基因。对24个全长基因完成了功能注释,并进一步开展了生物信息学分析,发现有6个应激反应基因和3个根发育相关基因。

For understanding the gene expression information in roots of trifoliate orange(Poncirus trifoliata),an excellent citrus rootstock,a full-length c DNA library was successfully constructed with the root material. According to the quality analysis results,the library reached 1.08 × 106 cfu · m L-1 in capacity with a recombinant efficiency as high as 95%. One hundred and eighty-two expressed sequence tags(ESTs,Gen Bank No.:JK316116–JK316297)were obtained from 200 clones that were randomly selected from the library,and then subjected to further assay. Among 96 unigenes obtained from those ESTs,which included 12 contigs and 84 singletons,68 unigenes were homologous with the genes in Gen Bank database while 56 unigenes were homologous with the citrus genes published. These unigenes may have biology function such as binding,catalytic activity and so on,and may play important roles in biology process such as cellular process,metabolic process,response to stimulus and so forth. Twenty-four unigenes were annotated among the 36 genes with the open reading frame(ORF)and 9 genes might be related with stress responsing or root developing.