The dehydration-responsive element-binding factor （DREB） is a plant-specific family of transcription factors and plays an important role in plant response and adaptation to abiotic stress. In the present work, two h...The dehydration-responsive element-binding factor （DREB） is a plant-specific family of transcription factors and plays an important role in plant response and adaptation to abiotic stress. In the present work, two highly similar CBF/DREB l-like genes, designated as PhCBF4a and PhCBF4b, were identified from Populus hopeiensis. These two genes contain all conserved domains known to exist in other CBF/DREB1 genes. In the AP2 domain, there is only one different amino acid residue between PhCBF4a and PhCBF4b, alanine or valine, a nonpolar amino acid, suggesting that PhCBF4a and PhCBF4b may have similar DNA binding ability. Their expression is induced by water-loss treatment and their expression patterns are similar. Moreover, with a genomic DNA as template, the presence of the same bands in PCR products as those in expression pattern analysis indicated that PhCBF4a and PhCBF4b exist in the genome ofP. hopeiensis. Their detailed functions are discussed and will need further study.展开更多
In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by ...In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by LD-PCR with the SMART cDNA Library Construction Kit, was in vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68×10^6 and 1.69×10^9 pfu·mL^-1 respectively. The combination ratio reached 98.8% and the average size of inserts was about 800 bp. In addition, the percentage of inserted fragments （〉400bp） was approximately 90%. The results indicate that a cDNA library has been successfully constructed.展开更多
基金the National Project of Science and Technology for the 11th Five-Year Plan in China (Grant No.2006BAD01A1502)
文摘The dehydration-responsive element-binding factor （DREB） is a plant-specific family of transcription factors and plays an important role in plant response and adaptation to abiotic stress. In the present work, two highly similar CBF/DREB l-like genes, designated as PhCBF4a and PhCBF4b, were identified from Populus hopeiensis. These two genes contain all conserved domains known to exist in other CBF/DREB1 genes. In the AP2 domain, there is only one different amino acid residue between PhCBF4a and PhCBF4b, alanine or valine, a nonpolar amino acid, suggesting that PhCBF4a and PhCBF4b may have similar DNA binding ability. Their expression is induced by water-loss treatment and their expression patterns are similar. Moreover, with a genomic DNA as template, the presence of the same bands in PCR products as those in expression pattern analysis indicated that PhCBF4a and PhCBF4b exist in the genome ofP. hopeiensis. Their detailed functions are discussed and will need further study.
文摘In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by LD-PCR with the SMART cDNA Library Construction Kit, was in vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68×10^6 and 1.69×10^9 pfu·mL^-1 respectively. The combination ratio reached 98.8% and the average size of inserts was about 800 bp. In addition, the percentage of inserted fragments （〉400bp） was approximately 90%. The results indicate that a cDNA library has been successfully constructed.
