Mutagenesis modified of Mx Gene from chicken and Identification of its antiviral specificity

Abstract:

[Objective] To study whether the antiviral specificity of chicken Mx protein is determined by an amino acid substitution at position 631. [Methods] We used PCR site-directed mutagenesis technique by which a single amino acid was reciprocally substituted G with A at position 2032bp of chicken Mx cDNA. Sequence analysis confirmed successful mutation from G to A at 2032bp of chicken Mx cDNA. The fragments amplified by PCR containing the mutation site were subcloned into a enteukaryotic expression vector. Then the recombinant vector was transfected into COS-Ⅰcell, Mx gene and Mx protein in the transfected COS-Ⅰcell were detected by RT-PCR and indirect fluorescence assay. [Results] The results showed that COS-Ⅰcell transfected the recombinant plasmid could stably express the Mx protein. The antivirus assay showed that Mx protein had characteristics of resistance to infection of Newcastle Disease Virus. [Conclusion] This study may provide a basis of the virus-resistant mechanism of Mx protein and production of virus-resistant transgenic chicken.