Department of General Biology, Vanderbilt University, Nashville, Tennessee 37235.
Plant Physiol. 1973 Jan;51(1):203-9. doi: 10.1104/pp.51.1.203.
Partially purified high molecular weight preparations of phytochrome, estimated to be close to 440,000 molecular weight based upon chromatography through a calibrated Bio-Gel P-300 column, were obtained from Garry and Newton oats (Avena Sativa L., cv. Garry and cv. Newton), rye (Secale cereale L., cv. Balbo), barley (Horedum vulgare L., cv. Harrison), and pea (Pisum sativum L., cv. Alaska) by a sequence of three chromatographic steps: brushite, diethylaminoethyl cellulose, and Bio-Gel P-300. No significant differences were observed between these preparations during purification or subsequent handling. In addition, a low molecular weight form of phytochrome was purified from Garry oats. Two specific antisera against a low molecular weight form of phytochrome (60,000 molecular weight) obtained from etiolated Garry oat seedlings are characterized and used to compare the phytochrome preparations. Double diffusion assays indicated antigenic identity between all preparations except that pea phytochrome yielded a spur when compared to oat phytochrome. Micro complement fixation assays yielded complete identity between Garry and Newton oat phytochrome, reduced activity with rye and barley phytochrome, and a complete lack of activity with pea phytochrome at the serum dilutions assayed. Immunoelectrophoretic assays indicated that all high molecular weight phytochrome preparations were homogeneous by this criterion and that there were only slight differences between the preparations in electrophoretic mobility. Large and small forms of phytochrome isolated from Garry oats were found to be very similar antigens when tested with the anti-small phytochrome sera, although the small form was observed to electrophorese at a much slower rate than the large.
部分纯化的高相对分子质量的光敏色素制剂，根据通过校准的 Bio-Gel P-300 柱层析的估计，接近 440,000 相对分子质量，从 Garry 和 Newton 燕麦（Avena Sativa L.，cv. Garry 和 cv. Newton）、黑麦（Secale cereale L.，cv. Balbo）、大麦（Horedum vulgare L.，cv. Harrison）和豌豆（Pisum sativum L.，cv. Alaska）中获得，通过三个色谱步骤的序列：磷酸氢钙、二乙基氨基乙基纤维素和 Bio-Gel P-300。在纯化或随后的处理过程中，这些制剂之间没有观察到显著差异。此外，还从 Garry 燕麦中纯化了一种低分子量形式的光敏色素。两种针对从小麦幼苗中获得的低分子量光敏色素（60,000 分子量）的特异性抗血清被表征并用于比较光敏色素制剂。双扩散试验表明，除豌豆光敏色素与燕麦光敏色素相比产生刺突外，所有制剂均具有抗原性。微量补体固定试验表明，Garry 和 Newton 燕麦光敏色素之间完全相同，与黑麦和大麦光敏色素相比活性降低，在测定的血清稀释度下与豌豆光敏色素完全没有活性。免疫电泳试验表明，所有高分子量光敏色素制剂均符合这一标准，并且在电泳迁移率方面，制剂之间只有微小差异。从小麦中分离出的大、小形式的光敏色素用抗小光敏色素血清进行测试时被发现是非常相似的抗原，尽管小形式被观察到电泳速度比大形式慢得多。
