| Construction and Characterization of cDNA Library from Water-Stressed Plantlets Regenerated in vitro of Populus hopeiensis |
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| 作者单位： | Wang Ze-liang Zhang zhi-yi* Lin Shan-zhi Lin Yuan-zhen Zhang Qian Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants，Ministry of Education，Beijing Forestry University，Beijing 100083，P. R. China |
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| 基金项目： | Supported by the National ＂863＂ Project （Grant No.2002AA2Z4011） |
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| 摘 要： | 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 n vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68×10i6 and 1.69×109 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 (> 400 bp) was approximately 90%. The results indicate that a cDNA library has been successfully constructed.
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| 关 键 词： | 干旱现象 森林保护 气候灾害 水压 DNA |
Construction and Characterization of cDNA Library from Water-Stressed Plantlets Regenerated in vitro of Populus hopeiensis |
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| Authors: | Wang Ze-liang Zhang zhi-yi Lin Shan-zhi Lin Yuan-zhen Zhang Qian |
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| Institution: | Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, Beijing 100083, P. R. China |
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| Abstract: | 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 1.68×106 and 1.69×109 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 (> 400 bp) was approximately 90%. The results indicate that a cDNA library has been successfully constructed. |
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| Keywords: | cDNA library Populus hopeiensis water-stressed plantlet characterization |
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