Posno M, van Vliet A, Groot G S
Nucleic Acids Res. 1986 Apr 25;14(8):3181-95. doi: 10.1093/nar/14.8.3181.
The nucleotide sequence of a Spirodela chloroplast DNA fragment, which directs the synthesis of a approximately 15 kD chloroplast ribosomal protein in an E. coli cell free system, has been determined. The deduced aminoacid sequence of the open reading frame shows extensive homology with E. coli ribosomal protein L16. Primer extension analysis, S1 nuclease mapping and nucleotide sequence analysis indicate that the chloroplast L16 gene (rpl16) is interrupted by a 1411 bp intron, which separates a short 5' exon from a large 3' exon. The shorter in vitro synthesized ribosomal protein results from an artificial initiation event at an internal ATG codon in the 3' exon. The sequences at the 5' and 3' splice sites of the intron are similar to consensus sequences described for other, class II intron containing, protein coding chloroplast genes. Northern hybridization experiments reveal 6 stable transcripts of rpl16 ranging from 500 b to greater than 4000 b. As determined by S1 nuclease mapping, the 3'-end of the smallest transcript maps exactly after the stem of a proposed termination signal. Finally, the implications of the finding of a cluster of several chloroplast ribosomal protein genes and possible polycistronic transcription of this chloroplast DNA region, are discussed in relation to the organization and expression of ribosomal protein genes found in the S10 operon of E. coli.
已确定了一个浮萍叶绿体DNA片段的核苷酸序列,该片段在大肠杆菌无细胞系统中指导合成一种约15kD的叶绿体核糖体蛋白。开放阅读框推导的氨基酸序列与大肠杆菌核糖体蛋白L16具有广泛的同源性。引物延伸分析、S1核酸酶图谱分析和核苷酸序列分析表明,叶绿体L16基因(rpl16)被一个1411bp的内含子中断,该内含子将一个短的5'外显子与一个大的3'外显子分开。较短的体外合成核糖体蛋白是由3'外显子内部的一个ATG密码子处的人工起始事件导致的。内含子5'和3'剪接位点的序列与其他含II类内含子的蛋白质编码叶绿体基因所描述的共有序列相似。Northern杂交实验揭示了rpl16的6种稳定转录本,大小从500b到大于4000b不等。通过S1核酸酶图谱分析确定,最小转录本的3'末端正好位于一个假定终止信号茎的后面。最后,结合大肠杆菌S10操纵子中核糖体蛋白基因的组织和表达,讨论了发现几个叶绿体核糖体蛋白基因簇以及该叶绿体DNA区域可能的多顺反子转录的意义。