In-gel expression and in situ immobilization of proteins for generation of three dimensional protein arrays in a hydrogel matrix.
In-gel expression and in situ immobilization of proteins for generation of three dimensional protein arrays in a hydrogel matrix.
复制标题
蛋白质的凝胶内表达和原位固定，用于在水凝胶基质中生成三维蛋白质阵列。
作者：
J. Byun;Kyung‐Ho Lee;Ka;Min;Dong
已经开发了一种用于在水凝胶基质上将DNA阵列直接转化为三维蛋白质阵列的方法。使用包埋有细菌蛋白质合成机器的琼脂糖凝胶作为DNA可编程表达凝胶基质，用于在DNA阵列上原位翻译基因。在将表达凝胶基质浇铸在DNA阵列上孵育后，在两个表面的界面处发生蛋白质合成，并且无细胞合成的蛋白质沉积在凝胶基质上相应的DNA斑点周围。通过用Ni-NTA部分修饰琼脂糖，使表达蛋白的扩散稀释最小化。该过程导致产生具有受限半径的局部蛋白质点。所开发的方法不仅简化了通常用于制备蛋白质阵列的程序，而且还提供了在阵列上加载更大量蛋白质的条件，同时在延长的时间段内保持其结构完整性和功能性。
A method has been developed for the direct conversion of DNA arrays into three dimensional protein arrays on a hydrogel matrix. An agarose gel embedded with bacterial protein synthesis machinery was used as the DNA-programmable expression gel matrix for the in situ translation of genes on a DNA array. Upon incubation of the expression gel matrix cast on a DNA array, protein synthesis took place at the interface of the two surfaces and the cell-free synthesized proteins were deposited on the gel matrix surrounding the corresponding DNA spots. Diffusional dilution of the expressed proteins was minimized by modifying the agarose with Ni-NTA moieties. This procedure resulted in the generation of localized protein spots with confined radii. The developed approach not only simplifies the procedures typically used for the preparation of protein arrays but it also provides conditions for the loading of higher amounts of proteins on the array while retaining their structural integrity and functionality over extended time periods.
影响因子：
16
作者：
Carlson, Erik D.;Gan, Rui;Hodgman, C. Eric;Jewett, Michael C.
通讯作者：
Jewett, Michael C.
影响因子：
15
作者：
Seefeld TH;Halpern AR;Corn RM
通讯作者：
Corn RM