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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Crystallization and identification of an assembly defect of recombinant antenna complexes produced in transgenic tobacco plants
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Crystallization and identification of an assembly defect of recombinant antenna complexes produced in transgenic tobacco plants

机译：转基因烟草植物中产生的重组天线复合物的结晶和组装缺陷的鉴定

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摘要

A chimeric Lhcb gene encoding light- harvesting chlorophyll a/b-binding protein (LHCII) was ex- pressed in transgenic tobacco plants. To separate native from recombinant LHCII, the protein was extended by six histidines at its C terminus. Recombinant LHCII was isolated by deter- gent-mediated monomerization of pure trimers followed by affinity-chromatography on Ni~2+-NTA-agarose (NTA is ni- trilotriacetic acid). Elution with imidazole yielded recombi- nant monomers that formed trimers readily after dilution of the detergent without further in vitro manipulations. LHCII subunits showed the typical chlorophyll a/b ratio at all steps of purification indicating no significant loss of pigments.
机译：在转基因烟草植株中表达了一个编码轻集叶绿素a / b结合蛋白（LHCII）的嵌合Lhcb基因。为了从重组LHCII中分离出天然蛋白，该蛋白在其C末端延伸了六个组氨酸。通过洗涤剂介导的纯三聚体单体化反应，然后在Ni〜2 + -NTA-琼脂糖上进行亲和层析（NTA为三三乙酸），分离重组LHCII。用咪唑洗脱得到的重组单体在稀释洗涤剂后无需进一步的体外操作即可轻松形成三聚体。 LHCII亚基在所有纯化步骤中均显示出典型的叶绿素a / b比，表明没有明显的色素损失。

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