The invention discloses a method for quickly constructing an IBV (
Avian Infectious Bronchitis Virus) reverse
genetic strain, and belongs to the technical field of
coronavirus reverse genetics. The constructing method comprises the following steps: quickly completing construction containing IBV genomic full-strength cDNA (Complementary
Deoxyribose Nucleic Acid) clone by taking a BAC (
Bacterial Artificial Chromosome) vector as a framework and applying an in-vitro
homologous recombination technology, directly transfecting cells by a constructed recombinant
plasmid, and transcribing in the cells, thus obtaining a transcript having
infectivity; completing
virus packaging; inoculating SPF (
Specific Pathogen Free) chick
embryo to a
mixed solution of the cells and a culture medium and passing from generation to generation, thus obtaining the IBV reverse
genetic strain. The constructing method disclosed by the invention has the advantages of simple operation and high positive
cloning efficiency, the obtained IBV reverse
genetic strain has passage stability, and an effective tool is provided for researching
pathogenesis of the
virus in vitro, developing a novel vaccine and the like; according to the method disclosed by the invention, transcription is carried out in the cells by utilizing a CMV (Cytomegalovirus)
promoter added on a 5' terminal, and the rescue efficiency of the
virus is greatly increased by utilizing an HDVR (
Hepatitis Delta Virus Ribozyme) sequence added on a 3' terminal.