Function Studies of a Rice Cysteine Protease Gene
Date Issued
2010
Date
2010
Author(s)
Yang, Hsiu-Chun
Abstract
The OsCPs (Os09g21370) gene from rice (Oryza sativa ssp japonica cv. Nipponbare) is expected to encode a cysteine protease, which has potential uses as cytotoxins for cell death when it triggered by constitutive promoter. Previously, three forms of this gene have been cloned as full-length and truncated (30 or 60 amino acids deleted from N-terminal). In this work, to determine and characterize the OsCPs gene’s functions, three forms of this gene were expressed constitutively or conditionally, by ligating the genes independently with CaMV-35S or PR-1a inducible promoter. The resulted six constructs were introduced into Arabidopsis, tobacco and rice plants by Agrobacterium-mediated transformation system. Analysis of the transgenic plants containing full-length and truncated OsCPs gene triggered by the CaMV-35S promoter indicate that neither full-length nor truncated form of the OsCPs gene leads to cell death. The expression of each construct was confirmed by RT-PCR technique with the mRNA templates extracted from the corresponding transgenic plants. Also, salicylic acid was applied to induce transgenic tobacco plants containing OsCPs gene triggered by the PR-1a promoter. Again, no lethal effect was observed and RT-PCR experiments demonstrated the expression of each form of the gene. Several ways were discussed to resolve the expression problems of OsCPs gene and the availability of the inducible system in the future.
Subjects
Cysteine protease
PCD
lethal gene
PR-1a promoter
Type
thesis
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