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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
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The purpose of this study was to characterize the T-DNAs introduced into the transgenic OsCK rice, as part of abiosafety evaluation. Choline Kinase (CK) gene is upregulated in the transgenic OsCK rice. We identified the insertion sites,flanking sequences, structures and sequences of the inserted T-DNAs. Based on the adaptor-ligation PCR, we found that the rightborder of the T-DNA was inserted at position no. 129971, on BAC clone OSJNBa0014J14 of chromosome 10. The flankingsequences of the left border region of the T-DNA (which was later identified as a region harboring a 1-kb long deleted sequence),could not be identified by various PCR-based trials. However, it was finally identified with whole-genome shotgun sequencing,using an Illumina sequencer. The result indicated that one of the T-DNAs was inserted into the CaMV 35S promoter region,whereas the other T-DNA was introduced at position 128947 on OSJNBa0014J14 clone, with an inverse orientation. During theinsertion process, a 1024-bp-long chromosome sequences flanked by the right border of the T-DNA region was deleted. A 370-bplong left border region and 199-bp long right border region corresponding to the matrix attachment region (MAR) sequences ofthe T-DNA were also deleted. Collectively, these results indicate that whole-genome shotgun sequencing is a useful tool to revealthe detailed sequences and structures of the introduced T-DNAs, especially in the case of multiple T-DNA insertions. Theexpenses incurred on genome sequencing can be easily compensated by minimizing the time and efforts invested in conventionalmolecular analyses.
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