中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

分离和纯化水稻(O.sativa)总信使RNA及其体外翻译

黄大年   

  1. 中国水稻研究所
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:1987-05-28 发布日期:1987-05-28

Isolation and Purification of mRNA and in vitro Translation for Rice(O.sativa)

Huang Danian   

  • Received:1900-01-01 Revised:1900-01-01 Online:1987-05-28 Published:1987-05-28

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摘要： 信使 RNA 是传递 DNA 遗传信息的工具,它也可以通过逆转录酶的催化合成互补 DNA(cDNA),用作分子杂交的探针,以分离特定的基因。本文选用10日龄水稻幼苗提取总 mRNA,52.6g 新鲜组织中获得6.5mg 总 RNA,根据 A260/A280=1.69,说明 RNA 中含极少量蛋白质。采用多聚尿苷酸-琼脂糖柱层析,从总的 RNA 中分离到88μg 总的 mRNA,免网织红细胞溶胶体外翻译体系产物表明,分离的 mRNA 具有生物功能。

关键词: 信使RNA, 多聚尿苷酸, 琼脂糖柱层析法, 体外翻译, 水稻

Abstract: The mRNA is a tool transferred genetic information for DNA and can be copied by reverse transcriptase to form a cDNA which can be used as a probe for molecular hybridization to isolate specific genes.At present selected young seedling of rice at 10 days to extract total mRNA.From 52.6mg fresh tissues got6.5mg total RNA.According toA260/A280=1.69,it indicated that a little amount protein was contained in RNA extracted.Applied poly(U)-sepharose column,88μg total mRNA was got from total RNA.The in vitro products for rabbit reticulocyte lysate translation system showed that isolated mRNA possessed biological function.

Key words: mRNA, Poly(U)-sepharose columm chromatography, in vitro translation, O.sativa