Phytochrome in photosynthetically competent plants: characterization by monoclonal antibodies. Progress report
Technical Report
·
OSTI ID:6097887
New monoclonal antibodies have been prepared to 124-kdalton phytochrome from etiolated oats, to phytochrome from etiolated peas, and to alkaline phosphatase. Simultaneously, progress has been made in improving methodologies for purification of phytochrome from green oats. Mice have been immunized with phytochrome electrophoretically purified from green oats. Methodologies needed to perform immunizations in vitro have been adapted to our purposes and screening protocols needed to identify antibodies to phytochrome from green oats are being refined. Forty-two rabbit antisera to phytochrome have been screened for their ability to immunoprecipitate phytochrome from green oats. The data are consistent with the hypothesis that green-oat phytochrome may be a minor component of the phytochrome isolated from etiolated oats. In addition, our panel of monoclonal antibodies has been screened for their ability to bind to phytochrome from green oats. Two that do so (oat-9 and oat-16) have been identified and characterized. A third (pea-25) has only recently been identified and is presently under intense investigation. Oat-9 and oat-16 apparently bind to the same domain on phytochrome, which is located near the site of chromophore attachment. They bind by ELISA to Pfr with greater affinity than to Pr. Pea-25 binds to phytochrome on immunoblots of sodium dodecyl sulfate, polyacrylamide gels. With this antibody it has been possible to determine that phytochrome from green oats appears to be identical in size to that obtained from etiolated oats. Data have also been obtained to indicate that a relatively small, but highly significant, proportion of the phytochrome obtained from green peas is immunochemically distinct from that obtained from etiolated peas. It thus appears that the observations made with green versus etiolated oats might represent a general phenomenon.
- Research Organization:
- Georgia Univ., Athens (USA). Dept. of Botany
- DOE Contract Number:
- AS09-81ER10925
- OSTI ID:
- 6097887
- Report Number(s):
- DOE/ER/10925-3; ON: DE85005904
- Country of Publication:
- United States
- Language:
- English
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