Amino Acid Substitutions in Matrix, Fusion and Hemagglutinin Proteins of Wild Measles Virus for Adaptation to Vero Cells

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Background: Wild-type measles virus (MV) is isolated in B95a but not in Vero cells. Through an adaptation process of wild-type MV to Vero cells, several amino acid substitutions were reported. Methods: Six strains were adapted to Vero cells and membrane (M), fusion (F) and hemagglutinin (H) genes were sequenced. Cell fusion was assessed and recombinant MVs were constructed, having wild-type H or M gene with or without mutations. Results: No F gene substitution was noted. Amino-acid substitutions at positions 481 from Asn to Tyr (N481Y) and 546 from Ser to Gly (S546G) were observed in the H protein. Glu at position 89 of the M protein was substituted for Gly (E89G) and two mutations were noted at positions 62 (S62R) and 83 (S83P) in M protein. Recombinant viruses with mutation(s) detected in Vero-adapted strains induced a cytopathic effect and grew well in Vero cells, but those with the wild type did not. Recombinant viruses with mutation(s) demonstrated lower viral growth in B95a cells. Conclusions: Substitutions of E89G, S62R and S83P of the M protein were newly observed through adaptation to Vero cells, besides the mutations described in previous reports, with varying adaptation for each strain. Copyright © 2011 S. Karger AG, Basel Received: January 27, 2010 Accepted after revision: July 26, 2010 Published online: January 13, 2011 Tetsuo Nakayama, MD Laboratory of Viral Infection I Kitasato Institute for Life Sciences, Kitasato University Shirokane 5-9-1, Minato-ku, Tokyo 108-8641 (Japan) Tel. +81 3 5791 6269, Fax +81 3 5791 6130, E-Mail tetsuo-n @ lisci.kitasato-u.ac.jp © 2011 S. Karger AG, Basel 0300–5526/11/0544–0217$38.00/0 Accessible online at: www.karger.com/int D ow nl oa de d by : 54 .1 91 .4 0. 80 9 /1 6/ 20 17 8 :2 4: 16 P M

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Amino Acid substitutions in matrix, fusion and hemagglutinin proteins of wild measles virus for adaptation to vero cells.

BACKGROUND Wild-type measles virus (MV) is isolated in B95a but not in Vero cells. Through an adaptation process of wild-type MV to Vero cells, several amino acid substitutions were reported. METHODS Six strains were adapted to Vero cells and membrane (M), fusion (F) and hemagglutinin (H) genes were sequenced. Cell fusion was assessed and recombinant MVs were constructed, having wild-type H o...

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تاریخ انتشار 2011