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Expression inE. coli of the cloned cDNA for the major antigen of foot and mouth disease virus Asia 1 63/72
Authors:V S V Suryanarayana  B U Rao  J D Padayatty
Institution:1. Department of Biochemistry, Indian Institute of Science, 560012, Bangalore, India
2. Indian Veterinary Research Institute, 560024, Hebbal, Bangalore, India
Abstract:Double stranded cDNA for the foot and mouth disease virus was prepared, restricted withBamH 1 or ligated to linkers withBamH 1 sticky ends and cloned inBamH 1 site in the expression vector, pU R222. The cDNA was also cloned at thePst 1 site in the same vector by the dC/dG tailing method. They were transferred intoE. coli to give colourless colonies in the presence of the dye, X-gal. Many of them showed positive signal on hybridization with32P-labelled viral RNA. The middleBamH1 fragment of the cDNA is known to carry the gene for the major antigen and some non-structural proteins. The clones carrying the recombinant DNA produced proteins which cross-reacted with the antibodies generated against the structural proteins of the virus in an enzyme linked immunosorbent assay, indicating that the cDNA of the major antigen is expressed in the cloned cell.
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