Gollmer I, Apel K
Eur J Biochem. 1983 Jun 15;133(2):309-13. doi: 10.1111/j.1432-1033.1983.tb07463.x.
Double-stranded cDNA was synthesized from polysomal poly(A)-containing RNA of illuminated barley plants and was inserted into the PstI site of the bacterial plasmid pBR322. Two different strategies were used for the screening of the bacterial colonies. Light-regulated sequences were detected by differential hybridization with cDNA of polyadenylated RNA from dark-grown and illuminated barley plants. For a second screening step partially purified mRNAs encoding the light-harvesting chlorophyll a/b protein were used for the synthesis of another cDNA probe. By using these procedures a cDNA clone was isolated which encodes a constitutive polypeptide of the light-harvesting chlorophyll a/b protein. This cloned cDNA has been used to assess the effect of phytochrome on the steady-state level of mRNA sequences encoding the light-harvesting chlorophyll a/b protein. The mRNA is almost undetectable in dark-grown plants. Following treatment with red light, the concentration of this mRNA sequence increases rapidly during the subsequent dark period. This red light effect can be reversed substantially by irradiation with far-red light. These results indicate that not only the amount of mRNA activity as shown previously, but also the steady-state level of mRNA sequences encoding the light-harvesting chlorophyll a/b is controlled by phytochrome.
从光照大麦植株的多聚核糖体含 poly(A) 的 RNA 合成双链 cDNA，并将其插入细菌质粒 pBR322 的 PstI 位点。采用了两种不同策略筛选细菌菌落。通过与黑暗生长和光照大麦植株的多聚腺苷酸化 RNA 的 cDNA 进行差异杂交来检测光调节序列。在第二步筛选中，使用部分纯化的编码捕光叶绿素 a/b 蛋白的 mRNA 来合成另一种 cDNA 探针。通过这些方法分离出一个 cDNA 克隆，它编码捕光叶绿素 a/b 蛋白的组成型多肽。这个克隆的 cDNA 已被用于评估光敏色素对编码捕光叶绿素 a/b 蛋白的 mRNA 序列稳态水平的影响。在黑暗生长的植株中几乎检测不到这种 mRNA。用红光处理后，在随后的黑暗期这种 mRNA 序列的浓度迅速增加。这种红光效应可以通过远红光照射而大大逆转。这些结果表明，不仅如先前所示的 mRNA 活性量，而且编码捕光叶绿素 a/b 的 mRNA 序列的稳态水平也受光敏色素控制。
