1995 Fiscal Year Final Research Report Summary
Formation of antibacterial medicinal plants by transformation using lysozyme gene
| Project/Area Number |
05557108
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
SHOYAMA Yukihiro Kyushu Univ.Fac.Pharm.Sci.Professor, 薬学部, 教授 (70037604)
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| Co-Investigator(Kenkyū-buntansha) |
MIKI Takeyoshi Kyushu Univ.Fac.Pharm.Sci.Associate Professor, 薬学部, 助教授 (40037586)
IMOTO Taiji Kyushu Univ.Fac.Pharm.Sci.Professor, 薬学部, 教授 (90038282)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Chichen egg lysozyme / Transformation / Antibacterial plant / Medicinal plant / 薬用植物 |
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| Research Abstract |
Nicotiana tabacum (1), Datura inoxia (2) and Rehmannia glutinosa (3) have heen co-cultivated with Agrobacterium tumefaciens strain LBA4404 which contained binary vector pBI121constructed using botn lysozyme and yeast invertase signal peptide carrying a regulatable CaMV35S promoter for effective secretion of expressed lysozyme to the intercellular spaces for antibacterial activity. The addition of 400 mg/l kanamycin was used for the selection of transformed callus and leaf segments.
(1) In the experiments of Nicotiana tabacum, the regenerated plantlets transformed with both lysozyme and signal-lysozyme genes secreted lysozyme which was determined by PCR using primer of lysozyme gene although the control plantlets did not secrete lysozyme.
(2) The highest line of GUS expression showed 45.6 times of GUS activity compared to untransformed callus. The transgenic nature was determined by the southern blot analysis and the histochemical assay. Lysozyme was not detected by the western blot analysis, although differences of lysozyme expression between the callus transformed with lysozyme gene and that transformed with signal-lysozyme chimeric gene was observed.
(3) Somatic embryos, leaf disks and shoot tips were used as infection segments. Regenerated plantlets were obtained both somatic embryo and leaf disk systems except that of shoot tip. However, the secretion of lysozyme has not been detected in all systems.
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