Abstract：Abstract: Fibrillisation dynamics under different pHs were analyzed for determination of the functions of spider silk protein modules in silk formation process. According to the complete MiSp gene sequence, R1R2CT fused with thioredoxin was expressed successfully via gene recombinant technology in BL21 (DE3), which resulted in a fusion protein as thioredoxin-thrombin cleavage site-R1R2CT. The expression level of recombinant proteins is about 15 mg/L LB medium. R1R2CT maintains soluble conformation and no positive ThT signals were detected under pH 7.5 and 6.5. Enhanced ThT signals were collected when pH decreased to 5.5, and during the following 3~5 h increased much more smoothly. After the first five hours, second swiftly sharp ThT signals appeared, and maintained to seven hours, after which the second relatively smooth increasement of ThT signals lasted all over the measurement time. The rate and latitude of R1R2CT ThT signal was higher than that of CT alone, and under pH 7.5, 6.5 and 5.5 no positive ThT signals were detected for R1R2 proteins indicated a quick fibrillisation process of R1R2 dependent on CT, and also CT is an essential part for spider silk formation. The fibrillation of R1R2CT to some extent supports the nucleation theory in some other light.
引用本文:
王 锋, 陈格飞, 孟 清. 蛛丝蛋白MiSp重组模块R1R2CT表达及其纤维化动力学研究[J]. 生命科学研究, 2015, 19(5): 410-414.
WANG Feng, CHEN Ge-Fei, MENG Qing. Recombinant Expression and Fibrillisation Analysis of MiSp R1R2CT Module. Life Science Research, 2015, 19(5): 410-414.
2015, Vol. 19 
