Effect of the Different Buffer on Electrophoretic Mobility Shift Assay (EMSA)

doi:10.7668/hbnxb.2009.06.036

Abstract

Abstract: The test of Electrophoretic Mobility Shift Assay is very simple in theory.But to successfully carry out gel shift experiments,it needs to optimize the parameters.Through a combination of T7 RNA polymerase and T7 Promoter for testing proper buffer was selected.The experimental results showed 5 × binding-Buffer 1 was more suitable for the test;TBE running2buffer was better than TAE and TGE buffer;0.× TBE buffer was the best concentration amang there electrophoresis buffer.

Key words: EMSA, Buffer, Optimize

CLC Number:

S665.1

ZHENG Hai-xia, WANG Yu-guo, FENG Zhi-fu. Effect of the Different Buffer on Electrophoretic Mobility Shift Assay (EMSA)[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(6): 178-180. doi: 10.7668/hbnxb.2009.06.036.

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References

[1] Fried M,Crothers D M.Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis[J].Nucleic Acids Res,1981,9(23):6505-6525.
[2] 梁国栋.最新分子生物学实验技术[M].北京:科学出版社,2001:4-9.
[3] Debomoy K,Lahiri,Yuan-Wen Gea.Electrophoretic mobility shift assay for the detection of specific DNA protein complex in nuclear extracts from the cultured cells and frozen autopsy human brain tissue[J].Brain Research Protocols,2000,5(3):257-265.
[4] Lance M Hellman,Michael G Fried.Electrophoretic mobility shift assay (EMSA) for detecting protein nucleic acid interactions[J].Nature Protocols,2007,2:1849-1861.
[5] Yue Li,Zhaozhao Jiang,Haixu Chen.A modified quantitative EMSA and its application in the study of RNA protein interactions[J].Journal of Biochemical and Biophysical Methods,2004,60(2):85-96.
[6] Woo A J,Dods J S,Susanto E,et al.A proteomics approach for the identification of DNA binding activities observed in the electrophoretic mobility shift assay[J].Mol Cell Proteomics,2002,1:472-478.
[7] 杨红梅,王磊.玉米转录因子zmCBF1的凝胶阻滞分析[J].生物技术通报,2005(6):81-83.
[8] 魏刚,雷娟,巩威,等.拟南芥QRAP2基因的克隆、表达、DNA结合能力及体外转录活性分析[J].科学通报,2005,50(17):1863-1868.
[9] 王海华,郝中娜,吴坤陆,等.水稻WRKY89中的亮氨酸拉链结构增强蛋白与W盒元件的相互作用[J].科学通报,2005,50(10):970-978.
[10] Karim Roder,Michael Schweizer.Running-buffer composition influences DNA-protein and protein-protein complexes detected by electrophoretic mobility-shift assay (EMSA)[J].Biotechnology and Applied Biochemistry,2002,33:209-214.
[11] Masashi Ono,Philip W Tucker,J Donald Capra.Ku is a general inhibitor of DNA-protein complex formation and transcription[J].Molecalar Immolunology,1996,33(9):787-796.

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