摘要
以25℃暗室中培养7d的阿拉斯加豌豆黄化苗为材料,采用改进的Yamamoto等方法,制备光敏色素粗提取液,再经DEAE-Sephacel,Brushite以及DEAE-Agarose柱层析得到初步提纯的光敏色素。然后再通过豌豆光敏色素抗体mAP1-Sepharose 4B亲和层析得到了提纯的豌豆114 kD光敏色素,经SDS-PAG电泳检测为一条带,并具有类似于纯净光敏色素的吸收光谱。
A 114 kD phytochrome preparation was obtained from etiolated seedlings of Pisum sativum cv. Alaska by purification. Phytochrome was extracted from etiolated pea seedlings (kept in darkness for 7 d at 25℃). The extract was purified by DEAE-sephacel-Brushite chromatography and DEAE-agarose column chromatography. After immunoaffinity chromatography (mAP1-Sepharose 4B), a 114 kD pea phytochrome was obtianed with an index of purification SAR (A_(667)/A_(280)) of 0. 73. SDS-PAG electrophoresis showed only one band. The purified phytochrome gave positive reaction with pea phytochrome monoclonal antibody (mAP13) and gave roughly the same absorption spectrum as that of native oat phytochrome.
基金
国家自然科学基金
关键词
豌豆
免疫亲和层析
光敏色素
Pisum sativum
immune affinity chromatography
monoclonal antibody
phytochrome
