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A Study on the Development of an Early Embryonic Gene of the Silkworm, Bombyx mori

누에 배형성기 초기 발현 유전자 개발 연구

  • Choi, Kwang-Ho (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Goo, Tae-Won (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kim, Seong-Ryul (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Park, Seung-Won (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kim, Sung-Wan (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA) ;
  • Kang, Seok-Woo (Sericultural & Apicultural Materials Division, National Academy of Agricultural Science, RDA)
  • 최광호 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 구태원 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 김성렬 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 박승원 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 김성완 (농촌진흥청 국립농업과학원 잠사양봉소재과) ;
  • 강석우 (농촌진흥청 국립농업과학원 잠사양봉소재과)
  • Received : 2012.04.18
  • Accepted : 2012.10.15
  • Published : 2012.12.30

Abstract

This study was aimed for a development of a useful gene promoter which has a transcript expressional specificity in the early embryonic period of the silkworm, Bombyx mori. To select a useful gene expressed in the early embryonic stage, we constructed and analyzed a PCR-base subtraction cDNA library. In subtractive hybridization analysis, we confirmed four clones as differently expressed genes(BmNanos-like, BmNanos-P, BmNanos-O, BmVasa mRNAs). Northern hybridization and real time PCR results reveled that the BmNanos-like gene promoter is suitable for the silkworm transgenic vector system. Further defined studies on molecular functions and biological roles of their promoters will give us well-fined information and its application.

본 연구는 누에 배자 발생 초기 특이 발현 유전자 프로모터를 개발하기 위한 연구의 일환으로 추진하였다. 누에 초기 및 후기 배자로 부터 분리한 mRNA를 사용하여 subtractive hybridization 분석법으로 누에 배자 발생 초기 특이 발현 유전자 4종을 선발할 수 있었다. 선발된 4종 유전자는 각각 BmNanos protein mRNA, BmNanos-P protein mRNA, BmNanos-O protein mRNA 및 BmVasa protein mRNA 유전자와 매우 높은 상동성을 보였다. 또한, 본 연구에서는 Northern hybridization 분석 및 real time PCR 분석을 통하여 배자 초기에 특이적으로 고발현하는 BmNanos-like 등 4개 선발 유전자의 발현 특성을 확인하였다. 이러한 결과는 추후 추진 할 누에 형질전환용 전이벡터의 효율성 제고를 위한 연구에 활용될 것으로 기대된다.

Keywords

References

  1. Yamanaka K, Inatomi S, Hanaoka M (1998) Cultivation characteristics of Isaria japonica. Mycoscience 39, 43-48. https://doi.org/10.1007/BF02461577
  2. Handler AM (2001) A current perspective on insect gene transformation. Insect Biochem Mol Biol. 31, 111-128. https://doi.org/10.1016/S0965-1748(00)00159-4
  3. Imamura M, Nakai J, Inoue S, Guo XQ, Kanda T, Tamura T (2003) Target gene Expression using the GAL4/UAS system in the silkworm Bombyx mori. Genetics 165, 1329-1340.
  4. Rika H, Tomita M, Yoshzato K (2006) The generation of germline transgenic silkworms for the production of biologically active recombinant fusion proteins of fibroin and human basic fibroblast growth factor. Biomaterials 27, 5715-5724. https://doi.org/10.1016/j.biomaterials.2006.07.028
  5. Satoshi Y, Zhu Z, Iaso K, Uchino K, Tamada Y, Tamura T, Asakura T (2007) Improving cell-adhesive properties of recombinant Bombyx mori silk by incorporation of collagen or fibronectin derived peptides produced by transgenic silkworms. Biomacromolecules 8, 3487-3492. https://doi.org/10.1021/bm700646f
  6. Tamura T, Thibert T, Royer C, Kanda T, Eappen A, Kamba M, Komoto N, Thomas JL, Mauchamp B, Chavancy G, Shirk P, Fraser M, Prudhomme JC, Couble P (2000). A piggyBac element- deribed vector efficiently promoters germ-line transformation in the silkworm, Bombyx mori L. Nat Biotechnol 18, 81-84. https://doi.org/10.1038/71978
  7. Tomita M (2011) Transgenic silkworms that weave recombinant proteins into silk cocoons. Biotechnol Lett 33, 645-654. https://doi.org/10.1007/s10529-010-0498-z
  8. White LD, Coares CJ, Atkinson PW, O'Brochta DA (1996) An eye color gene for the detection of transgenic non-drosophilid insects. Insect Biochem Mol Biol 26, 641-644. https://doi.org/10.1016/S0965-1748(96)00039-2