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 seedl...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.展开更多
以耐旱自交系邯郸177为材料,利用抑制性差减杂交技术(SSH),构建棉花苗期叶片的正向差减文库。挑取300个阳性克隆进行PCR验证,并对验证后的单克隆进行测序和分析,共获得284个有效序列。聚类后得到202条uniESTs序列,其中174条singlets,28...以耐旱自交系邯郸177为材料,利用抑制性差减杂交技术(SSH),构建棉花苗期叶片的正向差减文库。挑取300个阳性克隆进行PCR验证,并对验证后的单克隆进行测序和分析,共获得284个有效序列。聚类后得到202条uniESTs序列,其中174条singlets,28条contigs。经过BlastN分析,156个unigene可以在GenBank中找到同源序列,46个unigene未能找到同源匹配。经BlastX分析,40个unigene与未知功能蛋白或假定蛋白有较高相似性,116条unigene与已知功能蛋白有较高同源性。用KOBAS系统将33个unigene定位到55个Pathways中,其中P值小于0.5的Pathway有23条。初步分析发现,丙酮酸盐代谢(pyruvate metabolism)途径、乙醛酸和二羧酸代谢(glyoxylate and dicarboxylate metabolism)途径与棉花抗旱相关性较大。这些unigene基因涉及信号传导、能量代谢、蛋白质代谢、核酸代谢、光合作用及膜运输等代谢过程。发现了苹果酸合成酶基因(Ms1,001_B03;Ms2,003_E04)、苹果酸脱氢酶基因(Md1,001_C12;Md2,002_F01);NAC(001_C08)、锌指蛋白(zfp,003_C06)、BZR1/BES1(003_G04)等转录调节因子,以及翻译控制肿瘤蛋白基因(TCTP,002_C04)等耐旱相关基因。展开更多
文摘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.
文摘以耐旱自交系邯郸177为材料,利用抑制性差减杂交技术(SSH),构建棉花苗期叶片的正向差减文库。挑取300个阳性克隆进行PCR验证,并对验证后的单克隆进行测序和分析,共获得284个有效序列。聚类后得到202条uniESTs序列,其中174条singlets,28条contigs。经过BlastN分析,156个unigene可以在GenBank中找到同源序列,46个unigene未能找到同源匹配。经BlastX分析,40个unigene与未知功能蛋白或假定蛋白有较高相似性,116条unigene与已知功能蛋白有较高同源性。用KOBAS系统将33个unigene定位到55个Pathways中,其中P值小于0.5的Pathway有23条。初步分析发现,丙酮酸盐代谢(pyruvate metabolism)途径、乙醛酸和二羧酸代谢(glyoxylate and dicarboxylate metabolism)途径与棉花抗旱相关性较大。这些unigene基因涉及信号传导、能量代谢、蛋白质代谢、核酸代谢、光合作用及膜运输等代谢过程。发现了苹果酸合成酶基因(Ms1,001_B03;Ms2,003_E04)、苹果酸脱氢酶基因(Md1,001_C12;Md2,002_F01);NAC(001_C08)、锌指蛋白(zfp,003_C06)、BZR1/BES1(003_G04)等转录调节因子,以及翻译控制肿瘤蛋白基因(TCTP,002_C04)等耐旱相关基因。
