Schmidt R J, Myers A M, Gillham N W, Boynton J E
Department of Botany, Duke University, Durham, North Carolina 27706.
Mol Biol Evol. 1984 Jul;1(4):317-34. doi: 10.1093/oxfordjournals.molbev.a040320.
Polyclonal antibodies were elicited against seven of the 33 different proteins of the large subunit of the chloroplast ribosome from Chlamydomonas reinhardtii. Three of these proteins are synthesized in the chloroplast and four are made in the cytoplasm and imported. In western blots, six of the seven antisera are monospecific for their respective large subunit ribosomal proteins, and none of these antisera cross-reacted with any chloroplast small subunit proteins from C. reinhardtii. Antisera to the three chloroplast-synthesized ribosomal proteins cross-reacted with specific Escherichia coli large subunit proteins of comparable charge and molecular weight. Only one of the four antisera to the chloroplast ribosomal proteins synthesized in the cytoplasm cross-reacted with an E. coli large subunit protein. None of the antisera cross-reacted with any E. coli small subunit proteins. On the assumption of a procaryotic, endosymbiotic origin for the chloroplast, those chloroplast ribosomal proteins still synthesized within the organelle appear to have retained more antigenic sites in common with E. coli ribosomal proteins than have those which are now the products of cytoplasmic protein synthesis. Antisera to this cytoplasmically synthesized group of chloroplast ribosomal proteins did not recognize any antigenic sites among C. reinhardtii cytoplasmic ribosomal proteins, suggesting that the genes for the cytoplasmically synthesized chloroplast ribosomal proteins either are not derived from the cytoplasmic ribosomal protein genes or have evolved to a point where no antigenic similarities remain.
针对莱茵衣藻叶绿体核糖体大亚基的33种不同蛋白质中的7种产生了多克隆抗体。其中三种蛋白质在叶绿体中合成,四种在细胞质中合成并输入。在蛋白质免疫印迹中,七种抗血清中的六种对其各自的大亚基核糖体蛋白具有单特异性,并且这些抗血清均未与莱茵衣藻的任何叶绿体小亚基蛋白发生交叉反应。针对三种叶绿体合成的核糖体蛋白的抗血清与电荷和分子量相当的特定大肠杆菌大亚基蛋白发生交叉反应。针对在细胞质中合成的叶绿体核糖体蛋白的四种抗血清中只有一种与大肠杆菌大亚基蛋白发生交叉反应。没有一种抗血清与任何大肠杆菌小亚基蛋白发生交叉反应。基于叶绿体起源于原核内共生的假设,那些仍在细胞器内合成的叶绿体核糖体蛋白似乎比现在细胞质蛋白合成产物的那些蛋白与大肠杆菌核糖体蛋白保留了更多共同的抗原位点。针对这组在细胞质中合成的叶绿体核糖体蛋白的抗血清在莱茵衣藻细胞质核糖体蛋白中未识别出任何抗原位点,这表明在细胞质中合成的叶绿体核糖体蛋白的基因要么不是源自细胞质核糖体蛋白基因,要么已经进化到没有抗原相似性的程度。