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为研究沙冬青抗逆的分子机理,分离克隆其抗逆基因,以pBluescript II SK为载体构建了沙冬青(Ammopip-tanthus mongolicus)越冬叶片cDNA文库,并对文库的部分克隆进行了5’端随机测序,结果表明,该文库容量为2.16×105cfu,重组率为89.6%;插入片段的平均长度大于1 kb。随机测序得到542个EST,拼接成360个uniEST,经网上BlastN及BlastX分析,其中313个是已知功能的基因的标签,包括能量代谢、物质转化、细胞生长等生物过程,其中与抗逆相关的48条。
In order to study the molecular mechanism of the resistance of A. astigmatis, the resistance gene was isolated and cloned. The cDNA library of overwintering leaves of Ammopip-tanthus mongolicus was constructed by using pBluescript II SK as a vector. The partial clones of the library were randomly The results showed that the library capacity was 2.16 × 105 cfu and the recombination rate was 89.6%. The average length of the inserted fragment was greater than 1 kb. 542 ESTs were stochastically sequenced and 360 uniESTs were spliced. BlastN and BlastX were used to analyze the ESTs. Among them, 313 were ESTs with known functions, including biological processes such as energy metabolism, material transformation and cell growth. Among them, 48 of the