[Construction and screening of recombinant fowlpox virus expressing Eimeria tenella F2 hybrid strain SO7 gene].
OBJECTIVE
To construct the recombinant fowlpox virus expressing Eimeria tenella F2 hybrid strain SO7 gene.


METHODS
A recombinant expression plasmid pUTA-SO7 was constructed by inserting the SO7 gene of Eimeria tenella F2 hybrid strain into downstream of a hybrid poxvirus promoter which was flanked by the TK gene of fowlpox virus (FPV). The constructed pUTA-SO7 was firstly transfected into chicken embryo fibroblast cells(CEF) pre-infected with FPV strain 282E4 by using liposome, then the viruses resulted from the transfection were selected for 2 passages by culturing in CEF cells with MEM medium containing 40 mg/L 5-bromo-2-deoxy-uridine (BrdU). The selected viruses were plaque-purified in CEF cultured with MEM medium without BrdU. RESULTS Polymerase chain reaction (PCR), indirect immunofluorescence assay and Western blotting showed that S07 gene was expressed in recombinant fowlpox virus.


CONCLUSIONS
The recombinant FPV (rFPV) expressing the SO7 gene has been obtained.
Yang
Gui-lian
GL
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China.
Zhang
Xi-chen
XC
Zhao
Quan
Q
Li
Jian-hua
JH
Yin
Ji-gang
JG
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular / Q000379:methods; D004274:DNA, Recombinant; D016786:Eimeria tenella / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D005587:Fowlpox virus / Q000235:genetics*; D017125:Genes, Protozoan; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D004274:DNA, Recombinant; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins