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Identification, sequence characterization, and analysis of expression profiles of three novel CC chemokines from domestic duck (Anas platyrhynchos)
Authors:E Sreekumar  Avinash Premraj  D S Arathy  T J Rasool
Institution:(1) Animal Biotechnology Laboratory, Rajiv Gandhi Center for Biotechnology (RGCB), Thycaud PO, Trivandrum, Kerala, 695014, India;(2) Indian Veterinary Research Institute (IVRI), Mukteswar, Kumaon Campus, Nainital District, Uttaranchal, 263138, India
Abstract:Chemokines are low-molecular weight-chemotactic cytokines, which are involved in lymphocyte trafficking and migration of leucocytes to sites of injury, in immune surveillance and in healing process. They also play a role in pathogenesis of inflammatory diseases. Three novel CC chemokines were identified from domestic duck (Anas platyrhynchos) by screening of an enriched cDNA library constructed from mitogen-stimulated splenic mononuclear cells. Two of the clones (AB163 and AB330) had a very high nucleotide (both about 81%) and predicted amino acid level (71 and 76%, respectively) identity to the reported chicken macrophage inflammatory protein 1-beta (MIP-1beta; SCYA4) and regulated upon activation of normal T-cell expressed and secreted (RANTES; SCYA5) sequences. In phylogenetic analysis, these molecules clustered together with corresponding chemokines reported from other vertebrates. The third clone (AB187) had highest homology to chicken MIP-1beta (36% amino acid identity) and showed closer relation to a number of chemokines belonging to monocyte chemoattractant proteins and MIP-1agr chemokines. Expression of these molecules was upregulated upon mitogen stimulation of splenocytes as detected by semiquantitative RT-PCR. AB187 showed several fold increases (about 8.5 times) in the mRNA expression. Basal level expression of some of these chemokines was detected in both lymphoid and nonlymphoid tissues, including spleen, liver, lung, and bone marrow. Considering the importance of this animal species as a model for diseases such as chronic human hepatitis B, further studies will offer valuable insights into the role of these molecules in immunopathology of such diseases.The nucleotide sequences that are reported in this paper have been submitted to the NCBI Genbank database. Accession nos. AB163 (AY641435), AB187 (AY641436), and AB330 (AY641437).
Keywords:Subtraction  cDNA library  MIP-1beta  CCL4  CCL5
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