Characterization and membrane gene-based phylogenetic analysis of avian infectious bronchitis virus Chinese strain HH06.
A Chinese isolate of avian infectious bronchitis virus (IBV) designated HH06 was isolated from the kidney tissues of a chicken flock experiencing an outbreak of nephritis. In vivo pathogenicity of the IBV isolate HH06 was determined by inoculating specific pathogen-free (SPF) chickens. The clinical signs and related gross lesions of HH06 infected chickens were similar with those of the field-infected chickens. SPF embryonated eggs were inoculated with virus suspension for serial passage and their genomic RNA was extracted. RT-PCR technique was utilized to amplify the M gene sequence encoding membrane protein of IBV. Recombinant plasmid named T-vector-M was constructed via inserting the M gene into the TA cloning vector, pMD 18-T. The sequenced M gene and its deduced amino acid (aa) sequences were compared with the published sequences of reference strains. The M gene is of 687 bp in length encoding the M protein of 228 amino acids with a predicted molecular weight of 25.4 kDa. The sequences of the M gene and M protein share 83.9-97.9% and 83.6-96.5% homologous identities, respectively, compared with 29 IBV reference strains derived from different regions or countries, which revealed that there are still significant variations between strains. Furthermore, a phylogenetic tree based on these M DNA sequences was generated, and the tree topology suggests that some Chinese IBV strains may have a common ancestor; however, HH06 is a new local IBV isolate that is responsible for the field outbreak of nephritis.
Ren
Xiaofeng
X
College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai Street, 150030, Harbin, People's Republic of China. rxfemail@yahoo.com.cn
Yin
Jiechao
J
Ma
Dexing
D
Li
Guangxing
G
D000818:Animals; D002642:Chick Embryo; D002645:Chickens; D002681:China; D018352:Coronavirus Infections / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D000086862:Coronavirus M Proteins; D004196:Disease Outbreaks; D001351:Infectious bronchitis virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009393:Nephritis / Q000453:epidemiology / Q000662:veterinary / Q000821:virology; D010802:Phylogeny; D011201:Poultry Diseases / Q000453:epidemiology / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics*
D000086862:Coronavirus M Proteins; C000706230:M protein, Avian infectious bronchitis virus; D012367:RNA, Viral; D014763:Viral Matrix Proteins
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