Genomic profile of eGFP-tagged senecavirus A subjected to serial plaque-to-plaque transfers

文献类型： 外文期刊

第一作者： Chu, Huanhuan

作者： Chu, Huanhuan;Jin, Ningyi;Chu, Huanhuan;Wang, Jie;Liu, Fuxiao;Wang, Ling;Zhang, Youming;Jin, Ningyi;Li, Yan

作者机构：

关键词： Senecavirus A; eGFP; FP-FP transfers; Next-generation sequencing; Mutation; 3D protein; High-fidelity

期刊名称：MICROBIAL PATHOGENESIS （ 影响因子：3.8； 五年影响因子：4.0 ）

ISSN： 0882-4010

年卷期： 2024 年 191 卷

页码：

收录情况： SCI

摘要： Senecavirus A (SVA) belongs to the genus Senecavirus in the family Picornaviridae. This virus possesses a positivesense, single-stranded RNA genome, approximately 7200 nt in length, composed of a single 5 ' untranslated region, encoding region and 3 ' untranslated region. In this study, a recombinant SVA tagged with enhanced green fluorescent protein (eGFP) sequence, rSVA-eGFP, was rescued from its cDNA clone using reverse genetics. The passage-5 (P5) rSVA-eGFP was totally subjected to 55 rounds of consecutive fluorescent plaque-to-fluorescent plaque (FP-FP) transfers, and one extra common passaging in vitro. The P61 viral stock was analyzed by nextgeneration sequencing. The result showed ten single-nucleotide mutations (SNMs) in the rSVA-eGFP genome, including nine transitions and only one transversion. The P61 progeny still showed a complete eGFP sequence, indicating no occurrence of copy-choice recombination within the eGFP region during serial FP-FP transfers. In other words, this progeny was genetically deficient in the recombination of eGFP sequence (RES), namely, an RES-deficient strain. Out of ten SNMs, three were missense mutations, leading to single-amino acid mutations (SAAMs): F15V in L protein, A74T in VP2, and E53R in 3D protein. The E53R was predicted to be spatially adjacent to the RNA channel of 3D protein, perhaps involved in the emergence of RES-deficient strain. In conclusion, this study uncovered a global landscape of rSVA-eGFP genome after serial FP-FP transfers, and moreover shed light on a putative SAAM possibly related to the RES-deficient mechanism.

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