한국응용곤충학회지 (Korean journal of applied entomology)

  • 제61권1호
  • /
  • Pages.249-254
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  • 2022
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  • 1225-0171(pISSN)
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  • 2287-545X(eISSN)
한국응용곤충학회 (Korean Society of Applied Entomology)
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3D 프린팅 기술을 이용한 누에 알 미세주입 기술 개선

Improvement of Silkworm Egg Microinjection Using 3D Printing Technology

  • 정찬영 (국립농업과학원 농업생물부) ;
  • 이창훈 (국립농업과학원 농업생물부) ;
  • 석영식 (강원도 농산물원종장) ;
  • 용상엽 (강원도 농산물원종장) ;
  • 김성완 (국립농업과학원 농업생물부) ;
  • 김기영 (국립농업과학원 농업생물부) ;
  • 박종우 (국립농업과학원 농업생물부)
  • Jeong, Chan Young (Department of Agricultural Biology, National Institute of Agricultural Science) ;
  • Lee, Chang Hoon (Department of Agricultural Biology, National Institute of Agricultural Science) ;
  • Seok, Young-Seek (Gangwon-do Agricultural Product Registered Seed Station) ;
  • Yong, Sang Yeop (Gangwon-do Agricultural Product Registered Seed Station) ;
  • Kim, Seong-Wan (Department of Agricultural Biology, National Institute of Agricultural Science) ;
  • Kim, Kee Young (Department of Agricultural Biology, National Institute of Agricultural Science) ;
  • Park, Jong Woo (Department of Agricultural Biology, National Institute of Agricultural Science)
  • 투고 : 2021.12.29
  • 심사 : 2022.02.21
  • 발행 : 2022.03.01
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초록

오래전부터 산업곤충으로서 이용되어온 누에는 최근 바이오 신소재 생산을 위한 생체공장으로써 주목받고 있다. 소재생산을 위해서는 주로 형질전환 기술을 이용하게 되며, 이는 배아가 있는 알 속으로 목적 유전자를 삽입하는 마이크로인젝션(microinjection) 방식으로 이루어진다. 마이크로인젝션을 위해서는 알을 고정하는 과정이 필수적이며 시간 소모 및 피로도가 높은 작업이다. 따라서 본 연구에서는 시간 소모 및 피로도 개선을 통하여 형질전환 효율을 높이고자 3D 프린팅을 이용하여 알의 정렬 및 고정에 도움을 줄 수 있는 알 배열판(egg liner) 및 접착제 줄눈판(glue drawer)을 3DCADian 프로그램을 이용하여 모델링하고, Fusion 360를 이용하여 3차원 도면을 제작 후 프린팅하여 제작하였다. 제작된 두 도구를 이용하여 슬라이드 글라스에 알을 고정하고, 소요된 시간을 분석한 결과 도구를 사용하지 않았을 때에 비하여 2가지 도구를 이용했을 때 작업시간이 약18.6% 감소하였으며, 연구자의 작업 편의성을 향상시키고 마이크로인젝션을 위한 현미경 및 로봇 팔(manipulator) 조작을 유리하게 하였다. 따라서 알의 배열 수 또는 조작 편의성을 개선할 수 있는 추가적인 연구 및 개량이 이루어진다면, 알 배열판 및 접착제 줄눈판이 누에 형질전환 효율성 개선 및 다른 산업 곤충의 형질전환 연구에도 이용될 수 있을 것으로 판단된다.

Silkworms, which have for long been used as an insect resource for industrialization, have recently attracted attention as potential bio-factories for the production of novel biomaterials. In this regard, material production is typically achieved based on transformation technology, mediated via microinjection, in which a target gene is inserted into eggs containing an embryo. However, an essential step in the microinjection procedure is egg fixation, which can be a time-consuming and laborious task. Therefore, in this study, using the 3DCADian program, we adopted a 3D printing approach to model egg liners and glue drawers, which can contribute to facilitating egg alignment and fixation, thereby enhancing transformation efficiency by reducing time consumption and fatigue. After rendering using Fusion 360, the two supplementary tools were produced by printing with nylon resin (PA12) and Sinterit Lisa Pro. Subsequent analysis of the time required to fix eggs on glass slides using the two manufactured tools, revealed that the processing time was reduced by approximately 18.6% when the two tools were used compared with when these tools were not used. These innovations not only reduced fatigue but also contributed to more effective use of the microscope and manipulator for microinjection. Consequently, we believe that with additional research and refinement, the egg liner and glue drawer developed in this study could be used to enhance silkworm transformation efficiency and study similar transformation systems in other industrial insects.

키워드

과제정보

본 연구는 2021년도 농촌진흥청 공동연구사업(과제번호: PJ01622501) 및 국립농업과학원 전문연구원 과정 지원사업에 의해 이루어진 것임

참고문헌

  1. Coates, C.J., Jasinskiene, N., Miyashiro, L., James, A.A., 1998. Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti. Proc. Natl. Acad. Sci. U. S. A. 95, 3748-3751. https://doi.org/10.1073/pnas.95.7.3748
  2. Kang, P.D., Lee, S.U., Jung, I.Y., Shon, B.H., Kim, Y.S., Kim, K.Y., Kim, M.J., Hong, I.P., Lee, K.G., Park, K.Y., 2007. Breeding of new silkworm variety golden silk, a yellow cocoon color for spring rearing season. Korean J. Seric. Sci. 49, 14-17.
  3. Kim, T.K., Yong, H.I., Kim, Y.B., Kim, H.W., Choi, Y.S., 2019. Edible insects as a protein source: a review of public perception, processing technology, and research trends. Food Sci. Anim. Resour. 39, 521-540. https://doi.org/10.5851/kosfa.2019.e53
  4. Kumari, S.S., Subbarao, S.V., Misra, S., Murty, U.S., 2011. Screening strains of the mulberry silkworm, Bombyx mori, for thermotolerance. J. Insect Sci. 11, 1-14.
  5. Melgar-Lalanne, G., Hernandez-Alvarez, A.J., Salinas-Castro, A., 2019. Edible insects processing: traditional and innovative technologies. Compr. Rev. Food Sci. Food Saf. 18, 1166-1191. https://doi.org/10.1111/1541-4337.12463
  6. Okano, F., Satoh, M., Ido, T., Okamoto, N., Yamada, K., 2000. Production of canine IFN-gamma in silkworm by recombinant baculovirus and characterization of the product. J. Interferon Cytokine Res. 20, 1015-1022. https://doi.org/10.1089/10799900050198462
  7. Rubin, G.M., Spradling, A.C., 1982. Genetic transformation of Drosophila with transposable element vectors. Science 218, 348-353. https://doi.org/10.1126/science.6289436
  8. Tamura, T., Kuwabara, N., Uchino, K., Kobayashi, I., Kanda, T., 2007. An improved DNA injection method for silkworm eggs drastically increases the efficiency of producing transgenic silkworms. J. Insect Biotechnol. Sericol. 76, 155-159.
  9. Tamura, T., Thibert, C., Royer, C., Kanda, T., Abragam, E., Kamba, M., Komoto, N., Thomas, J.L., Mauchamp, B., Chavancy, G., Shirk, P., Fraser, M., Prudhomme, J.C., Couble, P., 2000. Germline transformation of the silkworm Bombyx mori L. using a piggyback transposon-derived vector. Nat. Biotechnol. 18, 81-84. https://doi.org/10.1038/71978
  10. Ueda, Y., Sakurai, T., Yanai, A., 1993. Homogeneous production of feline interferon in silkworm by replacing single amino acid code in signal peptide region in recombinant baculovirus and characterization of the product. J. Vet. Med. Sci. 55, 251-258. https://doi.org/10.1292/jvms.55.251