Nakatsuka Akira, Iwami Naoko, Matsumoto Shigehito, Itamura Hiroyuki, Yamagishi Masumi
Faculty of Life and Environmental Sciences, Shimane University, Japan.
Genes Genet Syst. 2002 Apr;77(2):131-6. doi: 10.1266/ggs.77.131.
We cloned and characterized Ty1-copia group retrotransposons in persimmon (Diospyros kaki Thunb.). Genomic DNA or methyl jasmonate (MJA)-treated cDNA were used as templates to amplify the reverse transcriptase region of Ty1-copia group retrotransposons. About 280 bp fragments were amplified and cloned, and 97 clones were sequenced. Forty-nine clones included frameshift or the stop codon, or both. Among 48 clones containing complete reading frames, 42 clones had unique nucleotide sequences. Alignment and phylogenetic analysis of putative amino acid sequences in the 42 clones indicated that these clones (named Tdk; retroTransposon in Diospyros kaki) fell into seven subgroups and six ungrouped sequences, indicating high sequence heterogeneity in Tdk clones. Phylogenetic analysis comparing unrelated plant species shows that some Tdk clones are more closely related to Ty1-copia group retrotransposons in the orders Solanales and Sapindales rather than to other Tdk clones. Southern blot analysis using Tdk2B, Tdk4c, Tdk6Ac, Tdk12K and Tdk13G clones as probes showed that persimmon and its related species, D. lotus, D. lotus var. glabba, D. oleifera, D. rhombifolia and D. virginiana, contained multiple Tdk-like sequences, indicating that homologous elements exist in other Diospyros species.
我们克隆并鉴定了柿树(Diospyros kaki Thunb.)中的Ty1-copia类逆转座子。以基因组DNA或茉莉酸甲酯(MJA)处理的cDNA为模板,扩增Ty1-copia类逆转座子的逆转录酶区域。扩增并克隆了约280 bp的片段,对97个克隆进行了测序。49个克隆包含移码或终止密码子,或两者都有。在48个包含完整阅读框的克隆中,42个克隆具有独特的核苷酸序列。对这42个克隆中假定氨基酸序列的比对和系统发育分析表明,这些克隆(命名为Tdk;柿树中的逆转座子)分为七个亚组和六个未分组序列,表明Tdk克隆中存在高度的序列异质性。对不相关植物物种进行的系统发育分析表明,一些Tdk克隆与茄目和无患子目植物中的Ty1-copia类逆转座子的亲缘关系比与其他Tdk克隆的更近。以Tdk2B、Tdk4c、Tdk6Ac、Tdk12K和Tdk13G克隆为探针进行的Southern杂交分析表明,柿树及其近缘物种君迁子、光叶君迁子、油柿、菱叶柿和弗吉尼亚柿含有多个Tdk样序列,表明其他柿属物种中存在同源元件。