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以强抗逆植物蒙古沙冬青(Ammopiptanthus mongolicus)为材料,用SMART技术构建了其在冷冻胁迫下的全长cD-NA文库。原始文库滴度为9.44×106pfu/ml,重组率为98.3%,插入片段长度在0.5～2.5kb,平均达到1kb。扩增文库滴度为1.98×1010pfu/ml。从原始文库中随机挑取480个阳性克隆进行序列分析,获得348个Unigene,其中有些是逆境应答相关基因和新基因。此外,以扩增文库为模板,经PCR方法克隆到读码框为1083bp的沙冬青类RD22基因。结果表明所建文库质量较高,可以满足后续研究的要求。
The full-length cDNA library of Ammopiptanthus mongolicus under freezing stress was constructed by using SMART technique. The titer of the original library was 9.44 × 106pfu / ml, the recombination rate was 98.3%, and the inserted fragment length was 0.5 ~ 2.5kb, averaging 1kb. The amplified library titer was 1.98 × 1010pfu / ml. 480 positive clones were randomly picked from the original library and sequenced. 348 Unigene were obtained, of which some were stress response related genes and new genes. In addition, the amplified library as a template, cloned by PCR method to read the 1083bp box of Astatine RD22 gene. The result shows that the quality of the constructed library can meet the requirements of the follow-up study.