디지털 미세유체를 이용한 미세녹조류 형질전환에서의 세포벽의 영향 분석

임도진; Do Jin Im

Clean Technology, Vol.21, No.2, 90-95, June, 2015

DOI10.7464/ksct.2015.21.2.090 Export Citation

디지털 미세유체를 이용한 미세녹조류 형질전환에서의 세포벽의 영향 분석

Effects of Cell Wall on the Transformation of Microalgae by a Digital Microfluidic System

임도진^†

국립부경대학교 화학공학과

Do Jin Im^†

Department of Chemical Engineering, Pukyong National University, 365 Sinseon-ro, Nam-gu, Busan 608-739, Korea

E-mail:

초록

디지털 미세유체 전기천공 시스템을 활용하여 미세녹조류에 대한 형질전환 실험을 통해 기존 상용화 장치 대비 높은 유전자 전달 효율과 세포 생존도를 확인하였다. 전기천공의 주요 파라미터인 인가전압 및 인가시간 변화를 주며 세포벽이 존재하는 세포종과 세포벽이 없는 세포종에 대한 비교 실험을 수행하였다. 이를 통해 식물 세포의 전기천공에서 세포벽의 역할은 단순히 유전체의 전달을 방해하는 부정적 요소로만 작용하지 않는 다는 사실을 확인하였다. 병렬화 및 온 칩 세포 배양등을 통해 제안된 디지털 전기천공 기술이 향후 새로운 청정 형질전환 방법으로써의 가능성도 확인하였다.

Digital microfluidic electroporation system was used for the transformation of microalgae and we have obtained higher transformation efficiency and viability than that of conventional method. Key parameters of electroporation such as pulse voltage, number, and duration time were systematically investigated for two different microalgal strains with and without cell wall. We have found that cell wall does not always have negative effects on the gene transformation of microalgae. Parallel processing of proposed digital microfluidic electroporation was demonstrated together with on chip culture of microalgae.

Keywords:Microalgae;Gene transformation;Droplet;Contact charging;Electrophoresis

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[1] Specht E, Miyake-Stoner S, Mayfield S, Biotechnol. Lett., 32(10), 1373 (2010)

[2] Guo SL, Zhao XQ, Tang Y, Wan C, Alam MA, Ho SH, Bai FW, Chang JS, J. Biotechnol., 163, 61 (2013)

[3] Gimpel JA, Specht EA, Georgianna DR, Mayfield SP, Curr. Opin. Chem. Biol., 17, 489 (2013)

[4] Basiouni S, Fuhrmann HJS, Biotechniques, 3, 1 (2012)

[5] Shimogawara K, Fujiwara S, Grossman A, Usuda H, Genetics, 148, 1821 (1998)

[6] Wang SN, Lee LJ, Biomicrofluidics, 7, 011301 (2013)

[7] Im DJ, Korean J. Chem. Eng., 32(6), 1001 (2015)

[8] Im DJ, Clean Technol., 20(4), 354 (2014)

[9] Im DJ, Noh J, Moon D, Kang IS, Anal. Chem., 83, 5168 (2011)

[10] Im DJ, Ahn MM, Yoo BS, Moon D, Lee DW, Kang IS, Langmuir, 28(32), 11656 (2012)

[11] Im DJ, Yoo BS, Ahn MM, Moon D, Kang IS, Anal. Chem., 85, 4038 (2013)

[12] Ahn MM, Im DJ, Kang IS, Analyst, 138, 7362 (2013)

[13] Lee DW, Im DJ, Kang IS, J. Phys. Chem. C, 117, 3426 (2013)

[14] Ahn MM, Im DJ, Kim JG, Lee DW, Kang IS, J. Phys. Chem. Lett., 5, 3021 (2014)