Definition: In this task, you're given a paragraph from the research paper and your task is to generate a suitable title for the research paper based on the given paper. Under 100 words is a good title length.
Input: MBP-tagged TRIM25 protein was expressed in E.coli Rosetta 2 (DE3) TM cells (Millipore). Transformed cells were grown in LB broth containing 0.2% glucose at 37 C. Then, 0.2 mM IPTG was added and the cells were incubated under shaking conditions for 18 h at 25 C, after which the cells were harvested by centrifugation. The pellet was resuspended in T1 buffer (20 mM Tris/Cl(pH 7.4), 1 mM EDTA, 1 mM sodium azide, and 1 mM DTT) and sonicated. After centrifugation, the supernatant was loaded onto a DEAE ion-exchange column (GE healthcare) and the flow throw was collected. Next, the pooled sample was applied to amylose resin (NEB). The column was washed with T2 buffer (T1 buffer + 200 mM NaCl) and eluted with T3 buffer (T2 buffer + 10 mM maltose). The purity of the proteins was analyzed by 10% SDS-PAGE. The final yield of MBP-TRIM25 was determined by the Bradford assay with BSA as the standard and the samples were stored at 80 C. His-tagged RIG-I was expressed in E. coli BL21(DE3) cells. The transformed cells were grown in LB broth at 37 C and then further incubated at 4 C for 3 h. Subsequently, 0.25 mM IPTG was added and the cells were incubated under shaking conditions for 18 h at 25 C, after which the cells were harvested by centrifugation. The pellet was resuspended in R1 buffer (50 mM HEPES; pH 7.4, 10% glycerol, 500 mM NaCl, 0.2% NP-40, 10 mM imidazole, 1 mM PMSF, 0.5 mg/ml lysozyme, and protease inhibitor cocktail) and sonicated. After centrifugation, the supernatant was applied to a Ni-NTA chelating agarose (Incospham). The column was washed with R2 buffer (50 mM HEPES; pH 7.4, 10% glycerol, 500 mM NaCl, 0.2% NP-40, and 50 mM imidazole) and the His-tagged RIG-I was eluted by increasing the imidazole concentration. The purest fractions were applied to a heparin-Sepharose column (GE healthcare), and the resin was washed with R3 buffer (50 mM Tris/Cl(pH 7.4), 10% glycerol, 0.2% NP-40, and 20 mM NaCl) and eluted with increasing concentrations of NaCl (up to 500 mM). The final yield of His-RIG-I was determined by the Bradford assay (Bio-Rad) with BSA as the standard. RNA Aptamers Specific to NS1 Protein of Influenza Virus Hye-Min Woo et al.
Output:
Molecules and Cells Supplementary Materials