Journal
PLANT MOLECULAR BIOLOGY REPORTER
Volume 27, Issue 3, Pages 388-399Publisher
SPRINGER
DOI: 10.1007/s11105-009-0114-3
Keywords
Heat-labile enterotoxin B; HN-neutralizing epitope; Newcastle disease virus; Transgenic tobacco; Chloroplasts
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Funding
- National Academy of Agricultural Science
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Synthetic enterotoxigenic Escherichia coli heat-labile enterotoxin B subunit-HN-neutralizing epitope fusion protein was expressed in E. coli and tobacco chloroplasts. Bacterial and chloroplastic recombinant LTB-HN-neutralizing epitope (LTB-HNE) fusion proteins showed the pentameric structures by sodium dodecyl sulfate polyacrylamide gel electrophoresis and a strong affinity for G(M1)-ganglioside. Bacterial and chloroplastic recombinant LTB-HNE was detected by Western blot analysis using polyclonal antibodies to HN-neutralizing epitope. Insertion of the gene encoding LTB-HNE protein into the chloroplast genomic DNA of spectinomycin-resistant plants was confirmed by polymerase chain reaction. The presence of the LTB-HNE specific transcript in the total RNA of transgenic plant leaves was verified by reverse transcriptase polymerase chain reaction. The highest level of expression of recombinant LTB-HNE fusion proteins in the leaves of transplastomic plants was about 0.5% of the total soluble protein. Growth rates, flowering, and seed setting were not affected, and the cassette for homologous integration and gene expression in the transplastomic T1 plant was subsequently inherited.
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