[Cloning and expression of gB gene of infectious laryngotracheitis virus in M. smegmatis].
Firstly, the complete gB gene of a domestic isolation stain were amplified by PCR, and the 2.6 kb gene fragment was obtained. Then the recombinant plasmid pY-alpha-gB was constructed by cloning PCR product into pY-alpha vector, and the shuttle expression plasmid pR-alpha-gB was constructed by cloning the hsp-alpha-gB gene into the downstream sequences of pRR3 vector. The recombinant plasmid was identified by restriction enzyme digestion and the sequence analysis, which was electrophoreted into M. smegmatis mc2 155. At last, the expressed gB proteins were successfully detected by ELISA and Western blot, which seems to be immunogenic crucially. The recombinant bacterial stain M. smegmatis mc2 155 (pR-alpha-gB)could protect SPF chick embryo from one lethal dose of ILTV.
Zheng
Hai-Zhou
HZ
College of Life Science, Hebei Normal University, Shijiazhuang 050016, China. zhenghaizhou@eyou.com
Yang
Hong
H
Bai
Jia-Ning
JN
Zhao
Bao-Hua
BH
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D015153:Blotting, Western; D002642:Chick Embryo; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D001352:Herpesvirus 1, Gallid / Q000235:genetics* / Q000276:immunology; D020102:Mycobacterium smegmatis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014614:Vaccines, Synthetic / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology
D000914:Antibodies, Viral; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C067381:GB protein, Infectious laryngotracheitis virus