[Cloning of glycoprotein B gene from strain Rispens of Marek's disease virus and construction of recombinant fowlpox virus].
Purified DNAs from Chicken Embryo Fibroblast (CEF) cultures infected with MDV strain Rispens were used as templates. Specific fragment with the size of about 2.9 kb was successfully amplified through Polymerase Chain Reaction(PCR) and identified to be gB gene of MDV by dot blot hybridization with a digoxigenin-labelled MDV gB specific oligonucleotide probe. The gB gene from strain Rispens was cloned into pUC19 and FPV insertion vector pFG1175-1 to construct plasmid pMGB and pFGBR1775-1 respectively. DOSPER liposome-mediated transfection with insertion vector DNA pFGBR1175-1 was performed on CEF monolayers infected with FPV 3-4 h earlier. Recombinant FPV was clone purified. Immunofluorescence Assay(IFA) showed that MDV gB gene had been expressed in FPV.
Xing
L
L
Department of Veterinary Medicine, Agricultural College, Yangzhou University, Yangzhou 225009.
Peng
D
D
Zhu
A
A
Liu
X
X
Zhang
R
R
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular; D005347:Fibroblasts / Q000821:virology; D005587:Fowlpox virus / Q000235:genetics*; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*
D000956:Antigens, Viral; D014759:Viral Envelope Proteins; C072811:glycoprotein B, Marek's disease virus