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Hua Y.,Southwest University for Nationalities | Hua Y.,Key Laboratory of Sichuan Institutes of Higher Learning | Hui D.Y.,Southwest University for Nationalities | Rong Z.H.,Southwest University for Nationalities | And 3 more authors.
Genetics and Molecular Research | Year: 2014

The full-length cDNA sequence of a novel expressed sequence tag (GenBank accession No. HQ184338) that was differentially expressed during Newcastle disease virus (NDV) infection in chickens was cloned from the chicken spleen by a rapid amplification of cDNA ends assay. This gene was further analyzed using bioinformatic methods and named grni. The full-length cDNA sequence was 1698 bp without introns, locating between 104,691,934 and 104,693,618 in galGal4 on chromosome 2. The open reading frame (ORF) contained 261 bp and encoded a deduced protein of 86 amino acid residues. Furthermore, the encoded protein contained two transmembrane regions without signal peptides, indicating that this protein is located in the mitochondrial membrane. Moreover, its homologous protein was not identified. Real-time polymerase chain reaction was used to detect the dynamic mRNA expression of this gene in the spleen, thymus, bursa of Fabricius, and trachea of NDV-infected chickens. Results suggested that the gene was involved in the transcriptional response of chicken to NDV infection. To obtain a fusion protein and prepare rabbit anti-serum, the predicted ORF of this gene was expressed in Escherichia coli. The expression of this gene at the protein level was further confirmed in the spleen, thymus, and bursa of Fabricius of NDV-infected chickens using Western blot analysis. In conclusion, a novel protein-coding gene named grni was successfully cloned and identified in chickens. Furthermore, this gene was found to be involved in the response of chickens to NDV infection. © FUNPEC-RP. Source

Zhang H.,Southwest University for Nationalities | Zhang H.,Key Laboratory of Sichuan Institutes of Higher Learning | Pi J.K.,Southwest University for Nationalities | Tang C.,Southwest University for Nationalities | And 5 more authors.
Avian Pathology | Year: 2012

Duck hepatitis A virus genotype C (DHAV-C), recognized recently, is one of the pathogens causing fatal duck viral hepatitis in ducklings, especially in Asia. To demonstrate the pathogenesis of the DHAV-C isolate, 3-day-old specific pathogen free ducklings were inoculated subcutaneously with a DHAV-C isolate and the clinical signs were observed. Virus distribution, histological and apoptotic morphological changes of various tissues were examined at different times post inoculation. The serial, characteristic changes included haemorrhage and swelling of the liver. Apoptotic cells and virus antigen staining were found in all of the tissues examined. Where more virus antigen staining was detected, there were more severe histopathological and apoptotic changes. The amount of virus antigen and the histological and apoptotic morphological changes agreed with each other and became increasingly severe with length of time after infection. Apoptotic cells were ubiquitously distributed, especially among lymphocytes, macrophages and monocytes in immune organs such as the bursa of Fabricius, thymus and spleen, and in liver, kidney and cerebral cells. Necrosis was also observed within 72 h post inoculation in all organs examined, except the cerebrum, and was characterized by cell swelling and collapsed plasma membrane. These results suggest that the recent outbreak of disease caused by DHAV-C virus is pantropic, causing apoptosis and necrosis of different organs. The apoptosis and necrosis caused by the DHAV-C field strain in this study is associated with pathogenesis and DHAV-C-induced lesions. © 2012 Copyright Houghton Trust Ltd. Source

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