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Comparative Studies of Wild-Type and Cold-Mutant (Temperature-Sensitive) Influenza Viruses: Polypeptide Synthesis by an Asian (H2N2) Strain and Its Cold-Adapted Variant

机译:野生型和突变型(温度敏感型)流感病毒的比较研究:亚洲(H2N2)菌株及其冷适应变异体的多肽合成

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摘要

The structure and replication of a cold-adapted, temperature-sensitive (TS) mutant of an Asian (H2N2) influenza virus was compared with that of its wild-type (WT) parent. Viruses were grown in a chicken kidney cell system, and at the nonpermissive temperature of 40 C, production of infectious TS virus was about 100,000-fold less than at 35 C, in contrast to WT virus. Major structural polypeptides of each virus grown at 35 C were similar, except that the hemagglutinin glycopolypeptide (HA) of the TS virions was slightly more heterogenous than that of WT virions. Synthesis of viral polypeptides was examined by sodium dodecyl sulfate acrylamide gel electrophoresis of pulse-labeled infected cells. This revealed a defect in the synthesis of TS viral hemagglutinin that was most pronounced at the nonpermissive temperature. Other TS viral polypeptides appeared to be synthesized normally at 40 C. A defect in the TS virus hemagglutinin was also indicated by serological studies that demonstrated that TS virus hemagglutinin had lost antigenic sites present on the WT virus. Thus, it is concluded that the virus mutant examined contains lesions in the hemagglutinin gene, although the possibility of additional unrecognized lesions is not excluded.
机译:比较了亚洲(H2N2)流感病毒的冷适应,温度敏感(TS)突变体的结构和复制与其野生型(WT)亲本的结构和复制。病毒在鸡肾细胞系统中生长,在40℃的非许可温度下,与WT病毒相比,传染性TS病毒的产生量比35℃少约100,000倍。在35 C时生长的每种病毒的主要结构多肽相似,只是TS病毒颗粒的血凝素糖多肽(HA)的异质性比WT病毒颗粒的异质性更高。病毒多肽的合成通过脉冲标记的感染细胞的十二烷基硫酸钠丙烯酰胺凝胶电泳检查。这揭示了TS病毒血凝素合成中的缺陷,该缺陷在非容许温度下最明显。其他TS病毒多肽似乎是在40°C正常合成的。血清病毒学研究还表明TS病毒血凝素存在缺陷,这些研究表明TS病毒血凝素失去了WT病毒上存在的抗原位点。因此,可以得出结论,尽管不排除其他无法识别的病变的可能性,但所检查的病毒突变体在血凝素基因中包含病变。

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