Abstract:To investigate the effects of mutation of 3D polymerase(3Dpol) from Enterovirus A71 (EV-A71), genomic cDNA was reverse transcribed from a parental EV-A71 and inserted into vector of TOPO-XL2-PCR to construct infectious clones. Two amino acid mutations designated N16S and I256V of 3Dpol were introduced into the 3Dpol encoding region as point mutation in vitro respectively. The viral genomic RNAs were transcribed by T7 polymerase and then transfected into RD cells, and the infectious parental and mutant viruses were rescued. Under normal temperate conditions (36.5 ℃), three rescued viruses possessed similar proliferation characteristics with viruse titers peaking at 72 hours-post-infection by viral infectivity titration and RNA content measurements. Temperature sensitivity test showed that all three rescued viruses had significant lower titers and RNA content at post-infection timepoints under higher temperature (39.5 ℃) condition compared to the normal temperature condition, indicative that the viruses were all sensitive to high temperature. Furthermore, compared to the parental virus, the proliferation of the viruses harboring the N16S or I256V mutant on 3Dpol were significantly inhibited under high temperature, suggesting that N16S or I256V mutation of 3Dpol of EV-A71 might affect the replication of the virus at high temperature.
朱颖, 翁育伟. 3D聚合酶突变对肠道病毒A组71型复制影响的研究[J]. 中国人兽共患病学报, 2021, 37(12): 1078-1083.
ZHU Ying, WENG Yu-wei. Effect of amino aicd substitution of 3D polymerase on viral replication in vitro of Enterovirus A71. Chinese Journal of Zoonoses, 2021, 37(12): 1078-1083.
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