Figure 2
Expression of Grttn in rLSDV-grttn infected FBT cells. (a) Immunostain of rLSDV-grttn-infected FBT cells incubated with RT specific sheep antibody, followed by peroxidase conjugated anti-sheep immunoglobulin antibody. (100X magnification). (b) Western blot analysis of cell lysates using sheep anti-RT and alkaline phosphatase conjugated anti-sheep. Lane 1, molecular weight markers; Lane 2, RT positive control protein standard; Lane 3, rMVA-grttn infected BHK-21 cells; Lane 4, rLSDV-grttn-infected FBT cells; Lane 5, LSDV infected FBT cells (c) Western blot analysis of cell lysates using sheep anti-p24 and alkaline phosphatase conjugated anti-sheep. Lane 1, molecular weight markers; Lane 2, Gag positive control protein standard; Lane 3, rLSDV-grttn infected FBT cells; Lane 4, LSDV infected FBT cells. (d) PCR analysis of rLSDV-grttn using primers designed to amplify a 1.4 kb fragment from the wild type LSDV and 1.2 kb fragment from the recombinant rLSDV-grttn. DNA products were subjected to 1% agarose gel electrophoresis. Lane 1, DNA molecular weight standards with relevant sizes indicated to the left; Lane 2, lysates of FBT cells; Lane 3, LSDV-infected FBT cells; Lane 4, rLSDV-grttn infected FBT cells.