期刊
JOURNAL OF PLANT BIOLOGY
卷 52, 期 3, 页码 244-250出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s12374-009-9023-0
关键词
Nicotiana benthamiana; Plastid transformation
资金
- Rural Development Administration [20070301034020]
- Crop Functional Genomics Center
- Korea Science and Engineering Foundation
- Korea Ministry of Education, Science, and Technology
- Korean Ministry of Marine Affairs and Fisheries
- Rural Development Administration (RDA), Republic of Korea [20070301034020] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Plastids from Nicotiana benthamiana were transformed with the vector for dicistronic expression of two genes-aminoglycoside 3'-adenyltransferase (aadA) and green fluorescent protein (gfp)-in the plastids of Nicotiana tabacum. Transplastomic shoots exhibited green fluorescence under UV light. Transformation efficiencies were similar between species. Although the border sequence (trnI and trnA) for homologous recombination to transform the plastid genome of N. benthamiana was identical to that sequence of N. tabacum, the exception was a 9-bp addition in the intron of trnI. This indicated that the N. tabacum sequence used as a border region for recombination was sufficient to insert the foreign gene into the target site between the trnI and trnA of N. benthamiana with similar efficiency. Southern blot analysis detected the presence of aadA and gfp between trnI and trnA in the plastid genome of N. benthamiana. Northern and western blot analyses revealed high expression of gfp in the plastids from petals and leaves. Our results suggest that the plastid transformation system established here is applicable to investigations of the interactions between plastid and nucleus in N. benthamiana.
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