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http://dx.doi.org/10.5010/JPB.2006.33.1.027

Characterization of Transgenic Lettuce (Lactuca sativa L.) Using a BL1 Gene Encoding Bromelain Isolated from Pneapple  

Jung, Yu-Jin (Department of Horticulture, Hankyong National University)
Kim, Gi-Hyun (Department of Horticulture, Hankyong National University)
Choi, Jang-Sun (Department of Horticulture, Hankyong National University)
Lee, Soon-Youl (Department of Genomic Engineering, Hankyong National University)
Nou, Il-Sup (Faculty of Plant Science and Production, Sunchon National University)
Park, Jin-Heui (Department of Horticulture, Hankyong National University)
Kang, Kwon-Kyoo (Department of Horticulture, Hankyong National University)
Publication Information
Journal of Plant Biotechnology / v.33, no.1, 2006 , pp. 27-32 More about this Journal
Abstract
To clarify the roles of bromelain in plants, we isolated BL1 gene encoding bromelain from pineapple stem tissues and sequenced. The full length cDNA is 933 bp and encodes a polypeptide of 311 amino acid residues. The cDNA is most similar 94% at the amino acid level to bromelain previously isolated from pineapple (BAA21929). Explants of Lactuca sativa were co-cultivated with Agrobacterium tume-faciences LBA 4404 strains containing nptII and BL1 gene for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 Ms basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were obtained T1 generation seeds with emasculation, and tested with PCR analysis using 35S promoter and BL1 specific primers whether BL1 gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot and endo protease activity showed that transcripts of BL1 gene were detected in transgenic lines. Theses results suggest that BL1 gene be successfully integrated and transcripted in the transgenic lettuce plants.
Keywords
Bromelain; overexpression; protease activity; transgenic Lettuce;
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