Studies on the structure and expression of chloroplast genes

将叶绿体DNA编码的基因定位到烟草叶绿体DNA物理图谱上，并克隆到质粒中。在他们中间。< α >基因的核苷酸序列。测定了atp酶< β >和<epsilon>亚基。测定了甘氨酸、精氨酸、丝氨酸和谷氨酰胺转移RNA基因的核苷酸序列。有趣的是，甘氨酸基因tRNA包含一个位于D茎上的691碱基对内含子。分析了DNA中一个大的反向重复和单拷贝区域之间的连接序列。从水稻幼叶中也分离到了叶绿体DNA。DNA通过几种限制性内切酶模式分析，估计DNA的大小为130 kbp。基于双酶切数据和Southern杂交，构建了sal1、Pst 1和Pvu 2的DNA物理图谱。利用烟草DNA片段作为探针，确定了水稻叶绿体DNA上几个与光合作用有关的基因的位置。由于水稻叶片叶绿体DNA的产量非常低。用Bam - HI和Pst - 1酶切产生的水稻叶绿体DNA片段克隆到质粒中。研究了Rubisco大亚基和32KD蛋白的基因表达。结果表明，这些蛋白质的mrna中有四分之一以不可翻译的形式存在于叶绿体中。这表明在翻译层面存在着调节机制。

The genes which are encoded by chloroplast DNA was located on tobacco chloroplast DNA physical map, and cloned into the plasmids. Among them. the nucleotide sequences of genes for <alpha> . <beta> and <epsilon> subunits of ATPase were determined. The nucleotide sequences of transfer RNA genes for glycine, arginine, serine and glutamine were also determined. It is interesting that tRNA for glycine gene contains a 691-base-pair intron located in the D stem. Sequence of the junctions between a large inverted repeat and single-copy regions in the DNA was analyzed.Chloroplast DNA was also isolated from young rice leaves. The DNA was analyzed by several restriction endonuclease patterns and the size of the DNA was estimated to be 130 kbp. A physical map of the DNA for Sal 1, Pst 1 and Pvu 2 has been constructed based on double digestion data and Southern hybridization.The location of several genes responsible for photosynthesis was also determined on rice chloroplast DNA using tobacco DNA fragments containing those genes as probes. Since the yield of chloroplast DNA from rice leaves is very low. rice chloroplast DNA fragments generated by the digestion with Bam HI and Pst 1 were cloned into the plasmids.Gene expression of the large subunit of Rubisco and 32KD protein was studied. It was shown that a quarter of mRNAs for these proteins exists as the untranslatable form in chloroplasts. This evidence suggests the existence of regulatory mechanism at the level of translation.