Patent ID: 8383334
Filing Date: 2013-02-26
Classification: C12N,C12Q,G01N

Abstract:
1. A method of screening libraries of variant proteins produced in plant leaves using a plant viral vector to identify a gene of interest, the method comprising: a) inoculating plant leaves with a library of viruses expressing variant genes; b) allowing sufficient time for a plurality of infected foci to form; c) harvesting a leaf with infected foci, the leaf having two opposing faces; d) sticking one face of the leaf to a sticky support material to immobilize the leaf and leaving the opposing face of the leaf exposed to form an exposed face; e) abrading the exposed face of the leaf with granular material to form an abraded face; f) placing the abraded face in contact with a blot membrane having a backing comprising blotting paper to form an assembly; g) placing the assembly into a vacuum seal bag; h) removing air and sealing the vacuum seal bag; i) removing the assembly and separating the blot membrane from the assembly; j) performing an assay on the separated blot membrane to identify an infected focus of interest; k) recovering virus of interest corresponding to the infected focus of interest; l) recovering nucleic acid from the virus of interest; and m) identifying the gene of interest from the nucleic acid.