Graduate Division of Biochemical Regulation, School of Agriculture, Nagoya University, 464-01, Nagoya, Japan.
Theor Appl Genet. 1993 Jun;86(5):579-84. doi: 10.1007/BF00838712.
In a previous study, we compared chloroplast DNAs (ctDNAs) from four species ofOryza and detected two independent deletions of DNA fragments in the ctDNAs (Kanno and Hirai 1992a). These deletions were genotype-specific variations. Since short direct-repeat sequences were detected at the borders of both deletions, the deletions were apparently the result of intramolecular recombination mediated by these direct-repeat sequences. In the present study, we examined whether or not this type of variation exists within a single species. Ishii et al. demonstrated three types of ctDNA inO. Sativa (1988), and the source of the variations that they identified seemed to be deletions. We determined the precise locations of the deletions and the sequences around them. As expected, our results showed that these variations were the results of deletions that were mediated by short direct-repeat sequences. While the deletions that had been found previously were located on spacer regions, those found in this study were located within open reading frames (ORFs). Northern hybridization analysis showed that one of the ORFs was-transcribed. In the case of this deletion, the amino acid sequence encoded by the C-terminal region of the ORF was altered and the short inverted-repeat sequences downstream of the ORF were deleted. In addition, there were other short inverted-repeat sequences downstream of the altered ORF.
在之前的一项研究中,我们比较了四个物种的叶绿体 DNA(ctDNA),并在 ctDNA 中检测到两个独立的 DNA 片段缺失(Kanno 和 Hirai 1992a)。这些缺失是基因型特异性的变异。由于在两个缺失的边界处检测到短的直接重复序列,因此这些缺失显然是这些直接重复序列介导的分子内重组的结果。在本研究中,我们检查了这种类型的变异是否存在于单一物种内。Ishii 等人在 1988 年证明了 O. Sativa 中的三种 ctDNA,他们确定的变异来源似乎是缺失。我们确定了缺失的精确位置及其周围的序列。正如预期的那样,我们的结果表明,这些变异是由短的直接重复序列介导的缺失的结果。虽然之前发现的缺失位于间隔区,但本研究中发现的缺失位于开放阅读框(ORF)内。Northern 杂交分析表明,其中一个 ORF 是转录的。在这种缺失的情况下,ORF 的 C 末端区域编码的氨基酸序列发生了改变,并且 ORF 下游的短反向重复序列被删除。此外,在改变的 ORF 下游还有其他短的反向重复序列。