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为了克隆和研究类胡萝卜素合成的一系列基因,采用TRIzol试剂提取枸杞叶片总RNA,然后利用PolyATtractmRNA分离系统分离mRNA,反转录合成第一链cDNA及第二链cDNA,以λ-ZAP为载体,体外包装后构建了滴度为2×107pfu/mL的枸杞叶片的cDNA文库。随机挑选10个克隆,经平板裂解酶切鉴定,插入序列的大小为0·5~3kb。该文库可以完全满足筛选或扩增基因的需要。
To clone and study a series of genes involved in the synthesis of carotenoids, TRIzol reagent was used to extract the total RNA from leaves of Lycium barbarum, and then the mRNA was isolated by PolyATtractmRNA isolation system. The first-strand cDNA and the second-strand cDNA were reverse transcribed to λ-ZAP The cDNA library of Lycium barbarum with a titer of 2 × 107pfu / mL was constructed after in vitro packaging. Ten clones were randomly selected and identified by restriction endonuclease digestion. The inserted sequence size was 0.5-3 kb. The library can fully meet the needs of screening or amplification of genes.