【求助】请问RNA聚合酶抑制的药物，使mRNA水平下降，蛋白..._RNA聚合酶_核酸酶类_分子类_知道

游客	收藏 \| 举报 2015-05-18 21:05 关注：238 回答：5 【求助】请问RNA聚合酶抑制的药物，使mRNA水平下降，蛋白... 已解决悬赏分：0 - 解决时间 2024-11-21 18:34 【求助】请问RNA聚合酶抑制的药物，使mRNA水平下降，蛋白水平不变正常吗

游客	举报 2015-05-22 07:42 How did you determine test gene mRNA levels? After 20 folds down, are levels of test gene mRNA sustained? Have you tried immunoprecipitation with S35 tagged proteins prepared from testing cells?

游客

举报 2015-05-19 03:00

mpark
How did you determine test gene mRNA levels? After 20 folds down, are levels of test gene mRNA sustained? Have you tried immunoprecipitation with S35 tagged proteins prepared from testing cells?

是用q-PCR,Trizol提取法。机器是Bio-rad的，20倍下降是给药最高浓度，用的2^(-△△CT)算法，24h，之后的时间点都没做，时间再长细胞毒性就比较显著了，细胞会皱缩减少。
S35标记蛋白没有试过，实验室没有条件做放射性试验，是说像Alexa488这样的标记蛋白观察蛋白分布的变化吗、
谢谢老师

游客

举报 2015-05-19 08:58

S35-methionine will be incorporated into peptides when protein synthesis takes place in side cells. By immunoprecipitation with antibody against protein tested in the cell which has been treated with the RNA polymerase inhibitor. In this way you can evaluate levels of tested protein synthesis.

游客	举报 2015-05-20 08:01 It is not always true that downregulation of mRNA would lead to declined levels of protein. It could stimulate protein synthesis by downregulating mRNA via inhibition of RNA polymerase.

游客	举报 2015-05-23 08:36 mpark It is not always true that downregulation of mRNA would lead to declined levels of protein. It could stimulate protein synthesis by downregulating mRNA via inhibition of RNA polymerase. 谢谢老师，S35实验室条件可能做不了，我继续查一下有没有RNA聚合酶被抑制使mRNA下降但蛋白合成不影响的情况，多谢老师

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【求助】请问RNA聚合酶抑制的药物，使mRNA水平下降，蛋白...