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KMID : 0903620040450020055
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Journal of the Korean Society for Horticultural Science
2004 Volume.45 No. 2 p.55 ~ p.59
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Methylation Pattern Analysis of Transgenic Tobacco Plants
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Lim, Chae Wan
Kim, Hyoung Seok/Oh, Yun Jung/Park, Young Doo
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Abstract
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Tabacco (Nicotiana tabacum cv. Havana SRI) was transfomed with Agrobacterium trmefaciens SBA4404 containing a rescue cloning vector (pRCV2). Southern hybridization data showed that the copy number of integrated genes varied from one copy to multiple copies. Transgenic plants confirmed with Southern hybridization were selfed and the T₁seeds from the transgenic lines with various copy numbers were tested in hygromycin medium to examine the propeny segregation ratio. The result showed that T-DNA was stably transferred to dight T₁lines. Polymerase chain reaction and Southern analyses were used to reconfirm the presence of hpt gene. Rescue cloning was performed to analyze methylation pattern of inserted T-DNA region. As a result, rescued plasmids were obtained from six lines and digested with AvaI AND ScaI. Cleaved-pattern appeared to methylated-pattern band or not. Particularly, the cleaved-pattern in line 14 revealed bands that were expected to methylated up to 355 promoter and hygromycin phosphotransferase coding region.
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