| Plant virus is a obligate parasites in plant cell or even the nucleus, it has caused serious damage to the agricultural production, people have been kooking for methods for controlling plant virus disease, but so far only found that some plants’ protein can inhibit the viral proteins, such as ribosome inactivating protein, but commercial production has not been evaluated. The study found a variety of anti-viral proteins associated with virion coat protein, understanding its mechanism contribute to the development of antiviral agents and breeding of antiviral transgenic crops, which are provided new solution strategies and control measures for the growing plant viral diseases.Extracted from edible mushroom Coprinus comatus in a basic glycoprotein Y3 to TMV(Tobacco mosaic virus, TMV) has inhibited, its mechanism may be in vitro inactivation of the Y3 to CP inhibits the synthesis of viral proteins, thereby reducing the rate of infection of the virus. To further verify the Y3 and CP is how the interaction, the test uses genetic engineering methods, respectively first obtain cDNA sequence of the protein Y3 and CP gene by cloning, Y3 gene cDNA sequence includes a 393 bp of ORF, the ORF speculated that may encode a polypeptide chain comprising 130 amino acids, CP gene cDNA sequence contains a 480 bp ORF, the presumed maximum ORF may encode a polypeptide chain containing 159 amino acids. Using ClustalX2.0 and DNAMAN6.0 software align cDNA sequences of obtained and published y3 and CP, Max identity, respectively 96% and 99%; Then the y3 gene cDNA sequence obtained connected to the carrier pGBKT7 to build bait vector, CP gene cDNA sequence connection with the carrier pGADT7 to build prey vector, which were severally transformed into yeast AH109 so that acquire fusion proteins, and finally pGBKT7-y3 bait vector and pGADT7-CP prey vector co-transformed into yeast AH109, using the yeast two-hybrid system detects proteinY3 and CP interaction,the results show the existence of protein Y3 and CP interaction, which has laid a good foundation for further study of its site of action and a clear mechanism of action. |
