Cloning and analysis of antiviral activity of a barramundi (Lates calcarifer) Mx gene☆
| Autor: | Shau-Chi Chi, Yu-Sheng Wu |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Myxovirus Resistance Proteins
Leucine zipper DNA Complementary Molecular Sequence Data Aquatic Science Biology Virus Replication Cell Line law.invention Rapid amplification of cDNA ends GTP-Binding Proteins law Complementary DNA Gene expression Birnaviridae Animals Cluster Analysis Environmental Chemistry Nodaviridae Amino Acid Sequence Cloning Molecular Gene Phylogeny Polymerase chain reaction DNA Primers Base Sequence RNA Sequence Analysis DNA General Medicine Virology Molecular biology Perciformes Open reading frame Gene Expression Regulation Sequence Alignment |
| Zdroj: | Fish & Shellfish Immunology. 23:97-108 |
| ISSN: | 1050-4648 |
| Popis: | We obtained a full-length cDNA clone for the Mx gene of barramundi (Lates calcarifer), using RACE (rapid amplification of cDNA ends) polymerase chain reaction (PCR) amplification of RNA extracted from a barramundi brain cell line cBB. The Mx cDNA of 2.2 kb contains an open reading frame (ORF) of 1875 nucleotides encoding a protein of 624 amino acids. The predicted barramundi Mx protein is 71.4 kDa and contains a tripartite guanosinetriphosphate (GTP)-binding motif at the amino terminal and a leucine zipper at the carboxyl terminal, characteristic of all known Mx proteins. Poly I:C-transfection induced the expression of Mx gene in cBB cells, and the induction level at 28 � C was higher than that at 20 � C. Moreover, Mx gene expression was also induced by viral infection, including fish nodavirus, birnavirus, and iridovirus. Among these, nodavirus was a stronger inducer than the other two viruses. Using an antiviral activity assay, we revealed that poly I:C-transfected cBB cells had antiviral activity against fish nodavirus and birnavirus, but not iridovirus. Furthermore, the replication of nodavirus and birnavirus could be restored after the expression of Mx genewas down-regulated by siRNA. Therefore, these results indicated that the expression of barramundi Mx gene was able to inhibit the proliferation of fish nodavirus and birnavirus. 2006 Elsevier Ltd. All rights reserved. |
| Databáze: | OpenAIRE |
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