Effects of genomic location of foreign gene in measles virus vector on gene expression and virus replication / 中华微生物学和免疫学杂志
Wenhao SU; Xiuxiu REN; Tingting ZHAO; Yinan WANG; Shishi LI; Qiufang HUANG; Xiaojie WANG; Xiaohuan ZHANG; Jiangbo WEI.
Chinese Journal of Microbiology and Immunology
; (12): 924-930, 2022.
Artículo
en Zh
| WPRIM | ID: wpr-995240
Objective:
To investigate the effects of genomic location of a foreign gene in Shanghai-191 strain measles virus (MV) vector on gene expression and virus replication.Methods:
The nucleotide sequence encoding S1 protein of SARS-CoV-2 was inserted at different positions in MV antigenome (the upstream of the N gene, between P and M genes, between H and L genes), and co-transfected into 293T cells with helper plasmids coding T7 RNA polymerase and N, P, and L proteins, respectively. The transfected cells were lysed and the supernatants were used to infected Vero cells to harvest recombinant viruses. S1 proteins expressed by the recombinant viruses were identified by RT-PCR, indirect immunofluorescence assay, Western blot and ELISA. Growth kinetics of the recombinant viruses were analyzed.Results:
Recombinant viruses were failed to be rescued when the S1 protein-coding sequence was cloned into the upstream of N gene. Two recombinant viruses, MV-M-S1 and MV-L-S1, were successfully rescued when cloning the S1 protein-coding sequence into the intergenic region between P and M genes, or H and L genes, and could express S1 protein. MV-M-S1 expressed more S1 protein than MV-L-S1, but the titer of MV-M-S1 was lower.Conclusions:
Inserting a foreign gene at different positions in the MV genome might have different effects on gene expression and virus replication. This study provided reference for the subsequent construction of MV vector.
Biblioteca responsable:
WPRO
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