Detailed Instructions: 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.
Q: Wild-type Middle East respiratory syndrome coronavirus (MERS-CoV) or I529T mutant MERS-CoV isolated from patients in South Korea (GenBank accession nos. KT029139.1 for wild type and KT868873.1 for I529T mutant) were cultured in a 24-well plate containing a monolayer of Vero E6 cells or 293T cells stably expressing CD26 (1). After 1 h incubation at 37C, viral supernatant was removed and cells were overlaid with 1 mL of 1% methylcellulose in Dulbecco modified Eagle medium, including 10% fetal bovine serum. Plates were incubated for 3 d at 37C, and then cells were fixed with 4% paraformaldehyde and 100% methanol. The MERS-CoV plaques were detected using rabbit anti-MERS-CoV N protein antibody (Sino Biologic Inc.) and goat anti-rabbit IgG secondary antibody conjugated with horseradish peroxidase (Invitrogen; https://www.thermofisher.com/us/en/home/brands/invitrogen.html). Viral plaques were visualized by incubation with 0.05% 33-diaminobenzidine tetrahydrochloride and 0.01% hydrogen peroxide in 50 mmol/L Tris-HCl (pH 8.0). Cellular layers were counterstained with trypan blue dye. Pseudotyped lentiviruses with wild type or mutant spikes of MERS-CoV were generated from 293T cells (Invitrogen) by cotransfection of human immunodeficiency virus backbone plasmids expressing firefly luciferase as previously described (1). We used the packaging plasmids, pLP1, pLP2, and pLP/VSV-G (Invitrogen) and pLVX-Luc-IRES-ZsGreen1 (Clontech; https://www.takarabio.com). For spike protein pseudotyping, condon-optimized cDNA of the
A:
Sequential Emergence and Wide Spread of Neutralization Escape Middle East Respiratory Syndrome Coronavirus Mutants, South Korea, 2015 Appendix Materials and Methods Middle East Respiratory Syndrome Coronavirus Culture and Plaque Assay