Silk G W, Wu M
Department of Biological Sciences, University of Maryland Baltimore County, Catonsville 21228.
Curr Genet. 1988 Aug;14(2):119-26. doi: 10.1007/BF00569335.
A plasmid library of chloroplast (Cp) DNA from Chlamydomonas reinhardtii was used to screen for transcripts which respond to light. A transcript of R03, a 1,300 bp EcoRI fragment, was identified as a message which accumulates in darkness. The transcribed region of R03 showed extensive sequence homology with the Escherichia coli elongation factor gene, tufA. A gene-specific probe was constructed. Northern blots were used to study the extent and kinetics of accumulation of this transcript in darkness and in the presence of antibiotic inhibitors of Cp ribosomes. For comparison, the effects of darkness and antibiotics on the steady state levels of psbA, rbcL, and 16S rRNA were also studied. We conclude that the tuf transcript shows the greatest increase in darkness and in the presence of antibiotics.
利用莱茵衣藻叶绿体(Cp)DNA的质粒文库筛选对光有反应的转录本。R03(一个1300 bp的EcoRI片段)的转录本被鉴定为在黑暗中积累的一种信使RNA。R03的转录区域与大肠杆菌延伸因子基因tufA显示出广泛的序列同源性。构建了一个基因特异性探针。利用Northern印迹法研究该转录本在黑暗中和存在叶绿体核糖体抗生素抑制剂时积累的程度和动力学。为作比较,还研究了黑暗和抗生素对psbA、rbcL和16S rRNA稳态水平的影响。我们得出结论,tuf转录本在黑暗中和存在抗生素时增加幅度最大。