Use of reverse genetics to develop a novel marker porcine reproductive and respiratory syndrome virus

Lin, Tao; Li, Xiangrui; Yao, Huochun; Wei, Zuzhang; Tan, Feiquan; Liu, Runxia; Sun, Lichang; Zhang, Rong; Li, Wenliang; Lu, Jiaqi; Tong, Guangzhi; Yuan, Sheng

doi:10.1007/s11262-012-0812-z

Cited by 3 publications

(1 citation statement)

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“…The nucleocapsid (N) protein is the most abundant protein in the PRRSV and plays a vital role in the viral life cycle, including regulating cytokine levels in the host, connecting with other viral proteins and combining with viral RNA [ 17 ]. As a protein with 123 or 128 amino acids, a mutation to an amino acid site may alter the functions of the N protein or affect viral replication ability and virulence [ 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

A novel amino acid site of N protein could affect the PRRSV-2 replication by regulating the viral RNA transcription

et al. 2022

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Background Finding the key amino acid sites that could affect viral biological properties or protein functions has always been a topic of substantial interest in virology. The nucleocapsid (N) protein is one of the principal proteins of the porcine reproductive and respiratory syndrome virus (PRRSV) and plays a vital role in the virus life cycle. The N protein has only 123 or 128 amino acids, some of key amino acid sites which could affect the protein functions or impair the viral biological characteristics have been identified. In this research, our objective was to find out whether there are other novel amino acid sites of the N protein can affect N protein functions or PRRSV-2 replication. Results In this study, we found mutated the serine78 and serine 99of the nucleocapsid (N) protein can reduce the N-induced expression of IL-10 mRNA; Then, by using reverse genetics system, we constructed and rescued the mutant viruses, namely, A78 and A99.The IFA result proved that the mutations did not affect the rescue of the PRRSV-2. However, the results of the multistep growth kinetics and qPCR assays indicated that, compared with the viral replication ability, the titres and gRNA levels of A78 were significantly decreased compared with the wild-type. Further study showed that a single amino acid change from serine to alanine at position 78 of the N protein could abrogates the level of viral genomic and subgenomic RNAs. It means the mutation could significant decrease the viral replication efficiency in vitro. Conclusions Our results suggest that the serine78 of N protein is a key site which could affect the N protein function and PRRSV replication ability.

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Section: Introductionmentioning

confidence: 99%