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以淹水处理 (submergence_treated ,ST)的玉米 (ZeamaysL .)幼苗根部cDNA为目标群体 ,未处理 (untreated ,UT)的玉米幼苗根部cDNA为对照群体 ,进行抑制差减杂交。用经过UT差减的STcDNA构建了一个含有大约 2 0 0 0个独立克隆的差减文库。对随机挑取的 40 8个克隆进行差异筛选 ,获得了 1 84个在ST中特异表达或表达增强的候选克隆。对其中 1 55个cDNA克隆测序并去除重复克隆后 ,共得到 95个差异表达的cDNA片段。GenBank中BLAST查询结果表明 :6个克隆为已知的玉米核苷酸序列 ;6 8个克隆与已知基因或EST序列部分区域的同源性为 6 0 %～90 % ;2 1个克隆在GenBank中无法查到对应的同源序列 ,可能代表了新基因 ,或者由于序列位于变异丰富的 3′端而无法查到与其他物种基因的同源性。
The root cDNA of maize (Zea mays L.) seedlings under submergence treatment (ST) was used as the target population, and untreated (UT) maize root cDNAs were used as the control group for suppression subtractive hybridization. A subtracted library containing approximately 200 independent clones was constructed with UT-subtracted STcDNA. A total of 408 randomly selected clones were screened by differential screening to obtain 1 84 candidate clones that were specifically expressed or enhanced in ST. A total of 95 differentially expressed cDNA fragments were obtained after sequencing a total of 115 cDNA clones and removing duplicate clones. The results of BLAST in GenBank showed that 6 clones were known maize nucleotide sequences. The homology of 6 8 clones to known gene or EST sequences was 60% ~ 90% GenBank can not find the corresponding homologous sequences, may represent a new gene, or because the sequence is located in the variable 3 ’end and can not be found with other species gene homology.