A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/...A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putative functions of ESTs were predicted by Blastn and Blastx.The 43 differentially expressed genes identified by subtractions were classified according to their putative functions generated by Blast analysis.Genetic functional analysis indicated that putative proteins encoded by these genes were related to diverse functions during organism development,which include biological regulation pathways such as transcription,signal transduction and programmed cell death,protein,nucleic acid and carbohydrate macromolecule degradation,the cell wall modification,primary and secondary metabolism and stress response.Two soybean transcription factors enhanced in SD conditions,GAMYB-binding protein and DNA binding protein RAV cDNAs that may be involved in SD soybean photoperiod response,had been isolated using 5'-and 3'-rapid amplification of cDNA ends(RACE)(Genbank Accession numbers DQ112540 and DQ147914).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A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putative functions of ESTs were predicted by Blastn and Blastx.The 43 differentially expressed genes identified by subtractions were classified according to their putative functions generated by Blast analysis.Genetic functional analysis indicated that putative proteins encoded by these genes were related to diverse functions during organism development,which include biological regulation pathways such as transcription,signal transduction and programmed cell death,protein,nucleic acid and carbohydrate macromolecule degradation,the cell wall modification,primary and secondary metabolism and stress response.Two soybean transcription factors enhanced in SD conditions,GAMYB-binding protein and DNA binding protein RAV cDNAs that may be involved in SD soybean photoperiod response,had been isolated using 5'-and 3'-rapid amplification of cDNA ends(RACE)(Genbank Accession numbers DQ112540 and DQ147914).
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。
