Construction of an infectious Marek's disease virus bacterial artificial chromosome and characterization of protection induced in chickens.
Marek's disease virus (MDV) is a highly oncogenic alphaherpesvirus that induces rapid-onset T-cell lymphoma in poultry. The complete genome of the avirulent vaccine strain MDV-814 was cloned as an infectious bacterial artificial chromosome (BAC) using an 8.8-kb fragment containing the self-designed selective marker guanosine phosphoriboxyl transferase. The recombinant virus MDV-814-BAC was generated by co-transfection of a BAC transfer vector and MDV-814 total DNA, and was purified by eight rounds of selective passaging. The infectivity of the BAC DNA clone was validated by MDV reconstitution from chicken embryo fibroblasts transfected with MDV-BAC DNA, which was extracted from electroporated Escherichia coli DH10B cells. In vitro, the BAC-derived virus had similar biological characteristics and growth kinetics as the wild-type parental and recombinant viruses, and chickens immunized with BAC derivatives by various delivery mechanisms acquired protection against virulent MDV challenge. Construction of this MDV-BAC may aid the development of recombinant vaccines-containing multiple antigens.
Cui
Hong-Yu
HY
Division of Avian Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China.
Wang
Yun-Feng
YF
Shi
Xing-Ming
XM
An
Tong-Qing
TQ
Tong
Guang-Zhi
GZ
Lan
De-Song
DS
He
Lai
L
Liu
Chang-Jun
CJ
Wang
Mei
M
D000818:Animals; D002645:Chickens / Q000276:immunology / Q000821:virology*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D005822:Genetic Vectors; D016679:Genome, Viral; D025922:Mardivirus / Q000235:genetics* / Q000254:growth & development / Q000276:immunology; D008380:Marek Disease / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D022501:Marek Disease Vaccines / Q000235:genetics* / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics
D004279:DNA, Viral; D022501:Marek Disease Vaccines; D019444:Vaccines, DNA