HIV-1 Tat蛋白和肽的比色测定

M. S. Radhi, A. R. Ruslinda, M. F. Fatin, S. S. B. Hashwan, M. Arshad, U. Hashim
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引用次数: 1

摘要

比色分析已被用于确定生物分子在溶液中的相互作用,这种相互作用可以通过分光光度计或肉眼观察到的颜色变化来指示。金纳米粒子(GNP)原本以粉红色溶液存在,但与盐溶液混合后会聚集并变为紫色。以其为基础溶液,与适体探针混合,加入NaCl盐溶液,在HIV-1 Tat蛋白靶点存在下,其颜色由粉红色变为紫色。由于其静电吸引、疏水吸收和共价结合的倾向,GNP能够吸附小的寡核苷酸。在本实验中,GNP吸附了覆盖其表面的适体探针,防止盐溶液与GNP相互作用。比色检测是通过观察靶物存在时适配体与蛋白质混合溶液的颜色变化和测量溶液波长的吸光度来证明其相互作用。由于颜色的变化,波长发生了变化。观察到GNP的波长值为522nm,在531.83nm处可以观察到该蛋白存在时的波长偏移,而在529.84nm处可以观察到该肽存在时的波长偏移。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Colorimetric assay of HIV-1 Tat protein and peptide
Colorimetric analysis has been used to determine the biomolecules interaction in a solution with that can be indicated from color changes that can be observed either by spectrophotometer or naked eyes. Gold nanoparticles (GNP) which originally exist as pink color solution will aggregate and turn to purple color when mixed with salt solution. It was used as the basic solution and it mixed with aptamer probe and when NaCl salt solution was added to the mixture, the color changes was observed from pink to purple in the presence of HIV-1 Tat protein target. GNP are capable of adsorbing small oligonucleotides due to their propensity for electrostatic attractions, hydrophobic absorption, and covalent binding. In this experiment, GNP adsorbed the aptamer probe covering its surface preventing salt solution from interacting with GNP. Colorimetric detection is use to prove the interaction that happened in the mixing solution of aptamer and protein by observing the color changes with the presence of target and measuring the absorbance of wavelength of the solution. The wavelength shift has been observed due to changes in color. The value of wavelength for GNP observed was 522nm and shifted in wavelength can be observed in the presence of tat protein at 531.83nm while for tat peptide at 529.84nm.
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