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A point mutation in the rhesus rotavirus VP4 protein generated through a rotavirus reverse genetics system attenuates biliary atresia in the murine model

Journal of Virology, . 2017;

s: Sujit K. Mohanty , Bryan Donnelly , Phylicia Dupree , Inna Lobeck , Sarah Mowery , Jaroslaw Meller , Monica McNeal , and Greg Tiao

Products/Services Used	Details	Operation
Peptide Synthesis	des 110 Synthetic peptides VSKLY, VSGLY, VSRLY, TRTRVSRLY and DGEA corresponding 111 to different regions on the VP4 protein of RRV were synthesized and purified by high112 performance liquid chromatography (HPLC) and kept lyophilized (GenScript, Piscataway, NJ). 113 Unrelated peptides (GHRP) of the same size and random sequence peptides with identical amino 114 acid (aa) compositions were synthesized for use as controls. Peptides were resuspended to 1 mM 115 in DMEM before use in replication assays.	Get A Quote

摘要

Recombinant KU 160 viruses carrying the RRV VP4s [KU-VP4(RRV)] were rescued from the 10th passage in MA104 161 cells by two plaque-to-plaque cloning steps in MA104 cells, as previously described (32). The KU-VP4(RRV) was back crossed to RT(VP4) 162 (RRV reassortant containing TUCH VP4) to obtain RRVrVP4 163 (RRV with a recombinant VP4). Therefore the viruses obtained from the plasmids pBacT7-VP4(RRV), pBacT7-VP4(RRVXbaI), and pBacT7-VP4(RRVR446G 164 ) were named RRVrVP4, RRVVP4-XbaI and RRVVP4-R446G 165 respectively.