title: Molecular cloning, expression and characterization of 100K gene of fowl adenovirus-4 for prevention and control of hydropericardium syndrome.
abstract: Fowl adenovirus-4 is an infectious agent causing Hydropericardium syndrome in chickens. Adenovirus are non-enveloped virions having linear, double stranded DNA. Viral genome codes for few structural and non structural proteins. 100K is an important non-structural viral protein. Open reading frame for coding sequence of 100K protein was cloned with oligo histidine tag and expressed in Escherichia coli as a fusion protein. Nucleotide sequence of the gene revealed that 100K gene of FAdV-4 has high homology (98%) with the respective gene of FAdV-10. Recombinant 100K protein was expressed in E. coli and purified by nickel affinity chromatography. Immunization of chickens with recombinant 100K protein elicited significant serum antibody titers. However challenge protection test revealed that 100K protein conferred little protection (40%) to the immunized chicken against pathogenic viral challenge. So it was concluded that 100K gene has 2397 bp length and recombinant 100K protein has molecular weight of 95 kDa. It was also found that the recombinant protein has little capacity to affect the immune response because in-spite of having an important role in intracellular transport & folding of viral capsid proteins during viral replication, it is not exposed on the surface of the virus at any stage.
authors.affiliation: Department of Pathobiology and Veterinary Sciences, University of Connecticut, Storrs, CT 06269, USA; Drug Discovery and Structural Biology Group, Health Biotechnology Division, NIBGE, Faisalabad, Pakistan. Electronic address: msalahuddin786@hotmail.com.
authors.forename: M S
authors.initials: MS
authors.lastname: Shah
authors.affiliation: Department of Zoology, Government College University, Faisalabad, Pakistan.
authors.forename: A
authors.initials: A
authors.lastname: Ashraf
authors.affiliation: Department of Pathobiology and Veterinary Sciences, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: M I
authors.initials: MI
authors.lastname: Khan
authors.affiliation: Drug Discovery and Structural Biology Group, Health Biotechnology Division, NIBGE, Faisalabad, Pakistan.
authors.forename: M
authors.initials: M
authors.lastname: Rahman
authors.affiliation: Vaccine Development Group, Animal Sciences Division, NIAB, Faisalabad, Pakistan.
authors.forename: M
authors.initials: M
authors.lastname: Habib
authors.affiliation: Drug Discovery and Structural Biology Group, Health Biotechnology Division, NIBGE, Faisalabad, Pakistan; Institute for Molecular Biology and Biotechnology (IMBB), The University of Lahore, Pakistan.
authors.forename: J A
authors.initials: JA
authors.lastname: Qureshi
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000276:immunology; D000257:Adenoviridae Infections / Q000235:genetics / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary / Q000821:virology; D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D002645:Chickens / Q000276:immunology / Q000821:virology; D003001:Cloning, Molecular; D005814:Genes, Viral / Q000276:immunology*; D007114:Immunization; D011201:Poultry Diseases / Q000235:genetics / Q000276:immunology / Q000517:prevention & control / Q000821:virology
chemical_list: D036022:Capsid Proteins
keywords: 100K; FAdV-4; Hydropericardium syndrome; Non structural protein; Phylogenetic analysis