[Mutagenesis modified of Mx gene from chicken and identification of its antiviral specificity].
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-I cell, Mx gene and Mx protein in the transfected COS-I cell were detected by RT-PCR and indirect fluorescence assay.


RESULTS
The results showed that COS- I cell 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.


CONCLUSIONS
This study may provide a basis of the virus-resistant mechanism of Mx protein and production of virus-resistant transgenic chicken.
Ni
Ligang
L
College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China. nlgangworld@gmail.com
Wu
Xiaowei
X
Cheng
Xumei
X
Chen
Hao
H
Chen
Qiang
Q
Song
Chengyi
C
Xu
Qi
Q
Li
Bichun
B
D019943:Amino Acid Substitution; D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D001483:Base Sequence; D019556:COS Cells; D002645:Chickens; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology*; D005822:Genetic Vectors / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D064547:Myxovirus Resistance Proteins; D009522:Newcastle disease virus / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000998:Antiviral Agents; D064547:Myxovirus Resistance Proteins; D019204:GTP-Binding Proteins