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. 2022 Aug 18;13:963009. doi: 10.3389/fimmu.2022.963009

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Copyright © 2022 Ruan, Feng, Cheng, Wang, Zhang, Wu, Yang, Tian, Ou, Sun, Zhang, Mao, Zhu, Jia, Chen, Liu, Zhao, Huang, Gao, Yu, Zhang and Pan

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation and identification of recombinant CHv with deletion of gC/gE genes. (A) Predicted and actual RFLP analysis of CHv and CHv-ΔgC/gE infectious clones. The obtained DNA was digested with BamH1 and electrophoresed in a 0.7% agarose gel. (line 1:BAC-CHv, line 2: BAC-CHv-ΔgC/gE) (B) Transfection of the plasmids CHv-ΔgC/gE -GS1783 into DEFs resulted in numerous fluorescent spots and cytopathies, the mutant virus CHv-ΔgC/gE were rescued. (C) PCR analysis of the gE expression of CHv-ΔgC/gE. (line 1: CHv-ΔgC/gE, line 2: Mock) (D) DEFs were infected (MOI = 0.01) with CHv-ΔgC/gE or CHv. At 24 h post infection (hpi), the levels of gC and gE proteins were determined with western blotting. (E) DEFs were infected (MOI = 0.1) with CHv-ΔgC/gE, CHv. Expression of gE and gI proteins was detected by indirect immunofluorescence at 36 hpi.