Abstract
Spinach is a long-day rosette plants, in which stem growth is mediated by gibberellins. It has been shown that two enzymatic steps, GA{sub 53}-oxidase and GA{sub 19}-oxidase, are controlled by light. To develop an understanding into this light regulation, purification of GA{sub 53}-oxidase has been undertaken. The original assay relied on the HPLC separation of the product and substrate, but was considered too slow for the development of a purification scheme. A TLC system was developed which in conjunction with improvements to the assay conditions was sensitive and gave rapid results. The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery. Monoclonal antibodies to the protein will be raised and used to further characterize the enzyme.
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Claims 3
Claim · model-written summary
Partial purification of GA53-oxidase from spinach yielded 1000-fold purification.
- Taxon
- Spinach
Spinacia oleraceae0 - Gene
- GA{sub 53}-oxidasee1
- Outcome
- purificatione1
- Value
- 1000-folde1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Spinach is a long-day rosette plants”
e1“The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery”
Claim · model-written summary
Partial purification of GA53-oxidase from spinach yielded 15% recovery.
- Taxon
- SpinachSpinacia oleraceae0
- Gene
- GA{sub 53}-oxidasee1
- Outcome
- recoverye1
- Value
- 15%e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Spinach is a long-day rosette plants”
e1“The partial purification of the GA{sub 53}-oxidase is achieved by a high speed centrifugation, 40-55% ammonium sulfate precipitation, an hydroxyapatite column, Sephadex G-100 column and an anion exchange FPLC column, Mono Q HR10/10, yielding 1000-fold purification and 15% recovery”
Claim · model-written summary
The TLC assay for GA53-oxidase was sensitive and gave rapid results, compared with the HPLC separation used originally.
- Taxon
- SpinachSpinacia oleraceae0
- Outcome
- rapid resultse3
- Qualifier
- sensitivee3
- Comparator
- HPLC separation of the product and substratee2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Spinach is a long-day rosette plants”
e2“The original assay relied on the HPLC separation of the product and substrate”
e3“A TLC system was developed which in conjunction with improvements to the assay conditions was sensitive and gave rapid results”