CAJ | 학술논문

以淹水处理(submergence-treated, ST)的玉米(Zea mays L.)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ST cDNA构建了一个含有大约2 000个独立克隆的差减文库.对随机挑取的408个克隆进行差异筛选,获得了184个在ST中特异表达或表达增强的候选克隆.对其中155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;68个克隆与已知基因或EST序列部分区域的同源性为60%～90%;21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.
이엄수처리(submergence-treated, ST)적옥미(Zea mays L.)유묘근부cDNA위목표군체,미처리(untreated, UT)적옥미유묘근부cDNA위대조군체,진행억제차감잡교.용경과UT차감적ST cDNA구건료일개함유대약2 000개독립극륭적차감문고.대수궤도취적408개극륭진행차이사선,획득료184개재ST중특이표체혹표체증강적후선극륭.대기중155개cDNA극륭측서병거제중복극륭후,공득도95개차이표체적cDNA편단.GenBank중BLAST사순결과표명:6개극륭위이지적옥미핵감산서렬;68개극륭여이지기인혹EST서렬부분구역적동원성위60%～90%;21개극륭재GenBank중무법사도대응적동원서렬,가능대표료신기인,혹자유우서렬위우변이봉부적3′단이무법사도여기타물충기인적동원성.
To investigate the expression profile of maize genes induced by submergence, a subtracted cDNA library of maize seedling roots was constructed using suppression subtractive hybridization (SSH). The cDNA of maize seedling roots treated with submergence (ST) was used as tester and what from untreated roots (UT) as driver. Products of the secondary PCR from the forward subtraction were cloned into T/A vector and transferred into Escherichia coli strain JM10B by electroporation. Four hundred and eight randomly chosen transformants carrying cDNA fragments were screened with PCR-Select Deferential Screening Kit. One hundred and eighty-four cDNA clones were identified as submergence specifically induced or highly expressed. After sequencing and removing redundant cDNAs, we got 95 submergence-induced cDNA clones. Of the 95 cDNA clones, 68 contain the regions with 60%-90% identity to their homolog in GenBank, 21 are expected to be novel genes, only 6 correspond to the published maize sequences.