Tsuchida Shirou, Kanashiki Takumi, Izumiya Shuhei, Ichikawa Takuya, Kurosawa Ryusuke, Hamaue Naoya, Aoki Takashi
Department of Molecular Biosciences School of Pharmaceutical Sciences Health Sciences University of Hokkaido Ishikari-Tobetsu Japan.
FEBS Open Bio. 2018 May 29;8(7):1176-1183. doi: 10.1002/2211-5463.12445. eCollection 2018 Jul.
The effect of the addition of sequential C-terminal tryptophan residues on the fluorescence intensity of GFP was investigated. Tandem repeats of six tryptophan residues markedly decreased fluorescence intensity. This phenomenon is likely to occur because of the inhibition of GFP folding, resulting in insolubility. Exploiting this phenomenon, we constructed a cloning vector that facilitates the identification of recombinant colonies of by the activation of GFP.
研究了连续添加C端色氨酸残基对绿色荧光蛋白(GFP)荧光强度的影响。六个色氨酸残基的串联重复显著降低了荧光强度。这种现象可能是由于GFP折叠受到抑制,导致不溶性。利用这一现象,我们构建了一种克隆载体,通过激活GFP来促进重组菌落的鉴定。