가뭄저항성벼(CaMsrB2-8)의 환경위해성 평가 : 3세대의 후대안정성 연구
Biosafety assessment for drought-tolerant transgenic rice (CaMsrB2-8) : Progeny stability study for three generations
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The purpose of this research is a molecular characterization of introduced T-DNA in transgenic rice T4~T6 generation lines (#8) harboring a pepper MsrB2 gene under the control of stress inducible Rab21 promoter, as a part of biosafety evaluation for drought-tolerant transgenic rice (CaMsrB2-8). We identified the structure and sequence of transformation vector of T-DNA and analyzed insertion sites, flanking sequences, and generational stability of inserted T-DNA in transgenic rice. The transformation vector was consisted of right border, a drought-tolerant CaMsrB2-8 gene unit (Rab21 promoter::CaMsrB2::PinII terminator), a selectable marker herbicide resistance unit (CaMV 35S promoter::bar::Nos terminator), and left border in sequential order. We confirmed that T-DNA was introduced at the position of 41,737,284 bp of chromosome No. 1. based on the adaptor-ligation PCR and whole genome sequence database. T-DNAs were stably inherited through the T4 to T6 generations, and also stable expression of bar gene from T-DNA was confirmed. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene (aadA) was not present in CaMsrB2-8 rice genome. These results will be filed to biosafety assessment document of CaMsrB2-8 rice.
Abstract
The purpose of this research is a molecular characterization of introduced T-DNA in transgenic rice T4~T6 generation lines (#8) harboring a pepper MsrB2 gene under the control of stress inducible Rab21 promoter, as a part of biosafety evaluation for drought-tolerant transgenic rice (CaMsrB2-8). We identified the structure and sequence of transformation vector of T-DNA and analyzed insertion sites, flanking sequences, and generational stability of inserted T-DNA in transgenic rice. The transformation vector was consisted of right border, a drought-tolerant CaMsrB2-8 gene unit (Rab21 promoter::CaMsrB2::PinII terminator), a selectable marker herbicide resistance unit (CaMV 35S promoter::bar::Nos terminator), and left border in sequential order. We confirmed that T-DNA was introduced at the position of 41,737,284 bp of chromosome No. 1. based on the adaptor-ligation PCR and whole genome sequence database. T-DNAs were stably inherited through the T4 to T6 generations, and also stable expression of bar gene from T-DNA was confirmed. It was also confirmed that the backbone DNA of transformation vector containing antibacterial gene (aadA) was not present in CaMsrB2-8 rice genome. These results will be filed to biosafety assessment document of CaMsrB2-8 rice.
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