Expression and purification of rice tungro coat protein (CP) genes for RTBV- and RTSV-specific antisera production
2004
Babb, G.M. | Dela Cruz, A.A. | Duque, M.J.C. | Sebastian, L.S.,Philippine Rice Research Inst., Maligaya, Science City of Munoz, Nueva Ecija (Philippines).
Maramara and others (2002) successfully cloned a coat protein gene of a local RTBV isolate using an innovative technique. This will be useful for specific antisera production for the tungro causal viruses and offers a good potential in genetics and breeding to confer CP-mediated resistance in transgenic rice. It has been difficult to biologically separate the RTBV from RTSV because of their complex double infection. The traditional procedure on preparation of RTBV and RTSV antigens is being hindered by the low individual virus concentrations in the plant. In addition, the purification method is very complicated with repeated use of toxic chemicals and ultra low to high-speed centrifugations. The protocol on CP expression and purification was enhanced and optimized to greatly reduce the inconvenience brought by the strenous traditional procedure. RTBV-CP2 was upscaled for RTBV-specific antisera production. Moreover, RTBV-CP1, -CP3 and RTSV-CP1, -CP2 and -CP3 genes were cloned and will be expressed in E. coli cells. These will serve as alternative sources of antigen for high-titer antisera production that will greatly facilitate tungro disease diagnosis in genetics and epidemiological research.
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