采用改进的酸酚法提取高质量的大豆叶片RNA,利用SMART思想和方法构建大豆叶片全长cDNA文库,直接以一级库液稀释液为模版进行PCR,快速克隆得到异黄酮代谢途径相关的5个基因。与传统的从DNA、RNA出发克隆基因,以及构建文库再进行基因筛选的克隆方法相比,该方法得到的基因均为全长基因,适用于快速、简便的进行多基因全长克隆。
High quality of total RNA from soybean leaves suitable for high efficient construction of cDNA library and gene cloning based on SMART strategy was isolated by the improved Acidic-Phenolic method. Five full length genes of isoflavonoid biosynthetic pathway were successfully and quickly amplified by PCR method with the original library cDNA dilution as templates. Comparing with traditional method of cloning genes from DNA or RNA, or isolating genes from cDNA plasmid hosted bacteria, this method can obtain full-length multi-gene more quickly and simply.
郝佳,马会勤,陈尚武. 利用SMART方法快速克隆异黄酮代谢途径多基因的研究[J]. 中国生物工程杂志, 2007, 27(6): 66-70.
. Cloning the multi-gene of isoflavonoid biosynthetic pathway with the SMART strategy. China Biotechnology, 2007, 27(6): 66-70.
https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2007/V27/I6/66
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