Enhancement of in vitro translation by gold nanoparticle--DNA conjugates.
Enhancement of in vitro translation by gold nanoparticle--DNA conjugates.
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DOI：
10.1021/nn100362m
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发表时间：
2010-05-25
期刊：
影响因子：
17.1
通讯作者：
Hamad-Schifferli K
中科院分区：
文献类型：
--
作者：
Park S;Hamad-Schifferli K
金纳米颗粒（AuNP）-DNA缀合物可以增强蛋白质的体外翻译。增强发生通过结合的非特异性吸附的相关分子和核糖体的AuNP-DNA和特异性结合的mRNA的兴趣。AuNP-DNA缀合物使网状裂解物混合物中荧光蛋白（mCherry，eGFP）的蛋白质产生增强65- 100%。凝胶电泳用于探测AuNP-DNA缀合物对翻译机器的非特异性吸附。确定非特异性吸附对于增强是关键的，并且如果消除非特异性吸附，则不发生表达增强。mRNA与AuNP表面上的DNA的相互作用影响增强的量，并且通过在RNase H存在下的表达来探测。这些结果表明，当AuNP上的DNA与mRNA形成不完全双链体时，发生更高的翻译增强。调整AuNP-DNA缀合物的非特异性吸附和特异性结合之间的平衡可以导致混合物中特定基因的翻译增强。
Gold nanoparticle (AuNP)-DNA conjugates can enhance in vitro translation of a protein. Enhancement occurs via a combination of non-specific adsorption of translation-related molecules and the ribosome to the AuNP-DNA and specific binding to the mRNA of interest. AuNP-DNA conjugates enhanced protein production of fluorescent proteins (mCherry, eGFP) in retic lysate mixes by 65–100%. Gel electrophoresis was used to probe non-specific adsorption of the AuNP-DNA conjugates to the translation machinery. It was determined that non-specific adsorption is critical for enhancement, and if it was eliminated, expression enhancement did not occur. The interaction of the mRNA with the DNA on the AuNP surface influenced the amount of enhancement, and was probed by expression in the presence of RNase H. These results suggest that higher translation enhancement occurs when the DNA on the AuNP forms an incomplete duplex with the mRNA. Tuning the balance between non-specific adsorption and specific binding of the AuNP-DNA conjugates could result in the translation enhancement of a specific gene in a mixture.
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