Abstract
The extraction and partial purification of phytochrome from light-grownAtrichum undulatum P. Beauv., a chlorophyllous moss, is described. Polyethyleneimine and salt fractionation followed by hydroxyapatite and Affi-gel-blue chromatography were used to separate phytochrome from chlorophyll, and to purify the pigment. All steps were performed in the presence of Triton X-100 which improved the yield by a factor of about three. The protein has a molecular weight some-what larger than that ofAvena phytochrome (124 kDa), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis. It cross-reacts with a monoclonal antibody against phytochrome from etiolated corn (Zea) and a polyclonal antibody against phytochrome from etiolated oat (Avena), and its photoreversibility is similar to that of phytochrome from greenAvena.
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Abbreviations
- EDTA:
-
ethylenediaminetetraacetic acid
- FMN:
-
flavinmononucleotide
- PMSF:
-
phenylmethylsulfonylfluoride
- Pr(Pfr):
-
red(far-red)-absorbing form of phytochrome
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
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Lindemann, P., Braslavsky, S.E., Hartmann, E. et al. Partial purification and initial characterization of phytochrome from the mossAtrichum undulatum P. Beauv. grown in the light. Planta 178, 436–442 (1989). https://doi.org/10.1007/BF00963812
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DOI: https://doi.org/10.1007/BF00963812