| 摘要 |
PURPOSE: A method for producing C4 recombinant protein of TYLCV and preparing antibody for the recombinant protein are provided to be used in a biological, molecular genetic, or immunological study. CONSTITUTION: A method for producing C4 recombinant protein of TYLCV(Tomato yellow leaf curl virus) comprises: a step of amplifying TYLCV-C4 gene through PCR, cloning the gene into pET28a vector, and transforming the vector into E.coli; a step of selecting transformed E.coli colonies and culturing; a step of adding IPTG to determine optimal culture condition; a step of expressing TYLCV-C4 recombinant protein; and purifying the TYLCV-C4 recombinant protein from the E.coli. A method for preparing monoclonal antibody of TYLCV-C4 recombinant protein comprises: a step of performing Ni-NTA resin chromatography under a denaturing condition of the TYLCV-C4 recombinant protein to elute TYLCV-C4-6X His fusion protein; a step of injecting the TYLCV-C4 recombinant protein into an experimental mouse three times; a step of colleting blood from the mouse and isolating antiserum; and a step of purifying IgG from the antiserum. |
