In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a b...In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on the beads. The double strand cDNA were released from magnetic beads by digestion with NotI, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA were directionally cloned intoλExcell vector. The results show that the primary titer of the cDNA library is 7.46×106 pfu per mL and the packaging efficiency reaches 1.47×107 recombinants per 靏 DNA. λDNA extracted from two clones of plaque were digested by EcoR I and NotI, both of the clones contained inserts larger than 900 bp. These results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed.展开更多
Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, ...Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, the gene response of Populus euphratica to salt stress is discussed. It is indicated that in response to salt treatment the transcription level for some genes of Populus euphratica increases by about 1.5 times and significant difference between the responses to osmotic stress and to ion stress has been observed in gene activity.展开更多
基金the National Natural Science Foundation of China (Grant No. 640650)
文摘In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on the beads. The double strand cDNA were released from magnetic beads by digestion with NotI, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA were directionally cloned intoλExcell vector. The results show that the primary titer of the cDNA library is 7.46×106 pfu per mL and the packaging efficiency reaches 1.47×107 recombinants per 靏 DNA. λDNA extracted from two clones of plaque were digested by EcoR I and NotI, both of the clones contained inserts larger than 900 bp. These results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed.
基金National Natural Science Foundation of China(Grant No.39830320)
文摘Through construction of a subtracted cDNA library and library screening, a number of salt-induced cDNA fragments have been cloned from Populus euphratica. Based on the results of DNA sequencing and Northern analysis, the gene response of Populus euphratica to salt stress is discussed. It is indicated that in response to salt treatment the transcription level for some genes of Populus euphratica increases by about 1.5 times and significant difference between the responses to osmotic stress and to ion stress has been observed in gene activity.
