Sequence comparison of MLEs of some lepidopteran insects inhabiting in east Asia isolated by PCR method using the inverted terminal repeat of Cecropia MLE and the analysis of transposase activity of MLE isolated from Attacus atlas of Yonaguni island by the same method.

• Project/Area Number: 14560048
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 蚕糸・昆虫利用学
• Research Institution: University of the Ryukyus
• Principal Investigator: NAKAJIMA Yumiko University of the Ryukyus, Center of Molecular Biosciences, Associate Professor, 遺伝子実験センター, 助教授 (70244340)
• Co-Investigator(Kenkyū-buntansha): KUSAKABE Takahiro Kyushu University Graduate School, Laboratory of Silkworm Science, Faculty of Agriculture, Assistant Professor, 大学院・農学研究院, 助手 (30253595)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2002: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: Cecropia-ITR-MLE / Inverted Terminal Repeat / Lepidoptera / Bombyx mori / Bombyx mandarina / Transposase : TP / BmTNML / Attacus atlas / Bombyx mandarina (クワコ) / トランスポゼース(TP:Transpposase) / 転移因子 / Mariner-like-element(MLE) / Transposase / Inverted Terminal Repeat(ITR)

Research Abstract

Mariner-like elements(MLEs) were isolated from several Asian insects(mainly lepidopteran insects) and invertebrates using the inverted terminal repeat(ITR) of the Hyalophora cecropia MLE as PCR primers. Around the discrete 1.3 kbp single bands from some specimens expected to contain the full-length of MLEs and various lengths of the fragments from other insects were detected. The full-length of MLEs isolated from the species inhabiting in relatively close regions around Japan were highly similar to each other classifying into the Cecropia MLEs sub family and some of them had the complete ORFs coding for transposase. This seem to indicate that the horizontal transfer of MLEs has taken place even among phylogenetically remote organisms recently and these MLEs might have the ability to transfer horizontally even now. Some of the clearly inactive MLEs making a complex isolated from Bombyx mori (n=28) and B.mandarina(n-27) were situated in other phylogenetic group than the horizontally group of Cecropia subfamily implies that this type of MLEs had inserted into the ancient genome of these species before the genomic segregation of B.mandarina between the Japanese/Korean types(n=27) and the Chinese/Korean types(n=28). One individual from China had a discrete 4.2-kbp band containing a BMCl and LlBm retrotransposon, which is a unique fragment detected in almost all of the silkworm races and strains(the MLE of this unit was named as BmTNML, Nakajima et al.,1999). Thus, the 4.2-kbp unit might be made spontaneously by crossing with B.mori after the segregation of B.mori genome.
The transposase region from the full length of mariner sequence isolated from Attacus atlas, which had the complete ORF for transposase, was cloned into the expression vector pENTR11. And the analyses of its transposition activity in the genome of E.coll and in the cultured cell of B.mori are still progressing now.

Research Products

• [Publications] Nakjima, Y., Fujimoto, H., Negishi, T., Hashido, K., Shiino, T., Tsuchida, K., Hidaka, M., Takada, N., Maekawa, H.: "Possible horizontal transfer of mariner-like sequences into some invertebrates including lepidopteran insects, a grasshopper and a coral."J.Insect Biotechnology and Sericology. 71. 109-121 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nakajima, Y., Nakamura, T., Banno, Y., Fujimoto, H., Hashido, K., Shiino, T., Tsuchida, K., Takada, N., Maekawa, H.: "Comparison of mariner-like elements among Bombyx mandarina individuals inhabiting east Asia in the light of the segregation of B.mori and B.mandarina."Int.J.Wild Silkmoth and Silk. 8. 57-64 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 中島裕美子, 佐藤良也: "文部科学教育通信NO77,6.9、最先端研究(水平伝播を含む活性型転移因子からみた生物ゲノムの多様性)"ジアース教育通信社. 28-29 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Nakajima, Y., Fujimoto, H., Negishi, T., Hashido, K., Shiino, T., Tsuchida, K., Hidaka, M., Takada, N., Maekawa., H.: "Possible horizontal transfer of mariner-like, sequences into some invertebrates including lepidopteran insects, a grasshopper and a coral."J.Insect Biotechnology and Sericology. 71. 109-121 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nakajima, Y., Nakamura, T., Banno, Y., Fujimoto, H., Hashido, K., Shiino, T., Tsuchida, K., Takada, N., Maekawa, H.: "Comparison of mariner-like elements among Bombyx mandarinaindividuals inhabiting east Asia in the light of the segregation of B.mori and B.mandarina."Int.J.Wild Silkmoth and Silk. 8. 57-64 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nakajima, Y., Nakamura, T., Banno, Y., Fujimoto, H., Hashido, K., Shiino, T., Tsuchida, K., Takada, N., Maekawa, H.: "Comparison of mariner-like elements among Bombyx mandarina individuals inhabiting east Asia in the light of the segregation of B.mori and B.mandarina."Int.J.Wild Silkmoth and Silk. 8. 57-64 (2003)
• [Publications] Nakajima, Y: "Possible horizontal transfer of mariner-like sequences into some invertebrates including lepidopteran insects, a grasshopper and a coral"J.Insect Biotechnology and Sericology. 71. 109-121 (2002)
• [Publications] Ohta, A.: "Cloning and characterization of testis-specific tektin in Bombyx mori"Comparatibe Biochemistry and Physiology Part B. (in press). (2003)
• [Publications] Lee, J: "Molecular characterization of a Heat Shock Cognate 70-4 promoter fro the silkworm, Bombyx mori"J.Insect Biotechnology and Sericology. 72. 33-39 (2003)