DOI number:10.16303/j.cnki.1005-4545.2021.12.03
Affiliation of Author(s):Yangzhou University
Journal:Chinese journal of veterinary science
Key Words:serotype 1 Fowl adenovirus; infectious clone; virus rescue; reverse genetics system
Abstract:The whole genome of the nonpathogenic serotype 1 fowl adenovirus (FAdV-1) JS 2017 strain previously isolated by our laboratory was extracted from infected LMH cells. Homologous sequences corresponding to the left terminal and right terminal of the genome (1 186 bp and 1 130 bp respectively) were amplified by PCR, and then cloned into the SuperCos-1 vector to generate a recombinant plasmid pCos-UD. Plasmid pCos-UD was linearized and co-transformed with the entire viral genome of JS2017 strain into E. coli BJ5183. By recombination between homologous sequences, the complete viral genomic DNA was inserted into the plasmid to generate an infectious clone pCos-FAdV1. The integrity of the inserted viral genome sequences in the infectious clones was confirmed by PCR assays targeting five different parts of the viral genome, and restriction enzyme digestion map analysis. The infectious clone pCos-FAdV1 was digested with Asc Ⅰ to release the FAdV-1 genomic DNA fragment from the vector and transfected into LMH cells by the liposome transfection method. The rescued virus was named cFAdV-1, and was shown to have similar replicability characteristics to the parent virus. In conclusion, the FAdV-1 infectious clone constructed in this study provides a technical platform of reverse genetic manipulation for further development of genetic engineering avian vaccines based on FAdV-1 vector.
Co-author:Yanlong Wang,Mengjiao Guo,Chengcheng Zhang,Zongyi Bo,Yongzhong Cao
First Author:Xiaorong Zhang
Indexed by:Research Atricle
Correspondence Author:Yantao Wu
Document Code:1005-4545（2021）12-2311-07
Discipline:Agricultural science
First-Level Discipline:Veterinary Medicine
Document Type:J
Volume:41
Issue:12
Page Number:2311-2317
Translation or Not:no
Date of Publication:2021-12-15