Park Won-Ji, You Sung-Hwan, Choi Hyoung-An, Chu Yeon-Jin, Kim Geun-Joong
Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Gwangju 500-757, Republic of Korea.
Department of Biological Sciences, College of Natural Sciences, Chonnam National University, Gwangju 500-757, Republic of Korea
Acta Biochim Biophys Sin (Shanghai). 2015 Jul;47(7):488-95. doi: 10.1093/abbs/gmv036. Epub 2015 May 20.
Specific tags with defined amino acid residues are widely used to purify or probe target proteins. Interestingly, the tagging system occasionally results in an increase of the recombinant protein expression in vivo. Here, we systematically examined this phenomenon using a poly-histidine (His)-tag fused to N- or C-terminal region of green, red, and blue fluorescent proteins by quantification and uneven distribution in cytoplasm of Escherichia coli. This effect was further supported by the distinct over-expression of several unrelated proteins, such as esterase, neopullulanase, and chloramphenicol acetyltransferase, tagging with the same tag. These results suggest that a poly-His-tag placed at N-terminal region can induce over-expression of recombinant protein via subcellular uneven distribution in vivo.
带有特定氨基酸残基的特异性标签被广泛用于纯化或探测目标蛋白。有趣的是，这种标签系统偶尔会导致体内重组蛋白表达增加。在此，我们通过对大肠杆菌细胞质中的绿色、红色和蓝色荧光蛋白的N端或C端区域融合多组氨酸（His）标签进行定量和不均匀分布分析，系统地研究了这一现象。几种不相关蛋白，如酯酶、新普鲁兰酶和氯霉素乙酰转移酶，用相同标签进行标记后出现明显的过表达，进一步支持了这一效应。这些结果表明，置于N端区域的多组氨酸标签可通过体内亚细胞不均匀分布诱导重组蛋白过表达。
