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인체 락토페린 생산 형질전환 고구마 개발

Development of transgenic sweet potato producing human lactoferrin

김재화|이영복|정원중|민성란|유장렬
한국식물생명공학회 (Korean Society for Plant Biotechnology)|2009.09|Journal of Plant Biotechnology|제36권 제3호|pp. 224-229

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참고문헌

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초록

면역강화와 같은 생리활성 기능을 가지는 인체 락토페린 cDNA가 들어있는 pLSM1 벡터를 이용하여 고구마배발생 캘러스에 particle bombardment 방법으로 형질전환을 수행하여 인체 락토페린 생산 고구마를 개발하였다. 선발배지에서 형성된 배발생 캘러스로부터 체세포배발생 과정을 거쳐 식물체로 재분화되었고 토양에서 정상적으로 생장하여 괴근을 형성하였다. 재분화된 식물체를대상으로 PCR과 Southern 분석을 실시한 결과 고구마의염색체 게놈에 인체 락토페린 유전자가 도입되었음을 확인하였다. 또한, northern blot 분석을 통해 락토페린 유전자의 발현을 확인하였으며 immunoblot 분석을 통해 고구마 괴근에서도 80 kDa 크기의 락토페린 단백질이 생산됨을 확인할 수 있었다. 본 연구결과 고구마는 치료용 인체단백질을 생산하는 숙주로서 매우 우수하다는 것을 입증하였다.

Abstract

Human lactoferrin is an iron-binding glycoproteinwith many biological activities, including the protectionagainst microbial and virus infection and stimulation of theimmune system. We introduced a human lactoferrin (hLf)cDNA under the control of 35S promoter into sweet potatoby particle bombardment. Transgenic plants were regeneratedvia somatic embryogenesis. Transgenic plants wereproduced typical tuberous roots in soil. PCR, Southern andnorthern analyses confirmed that the hLf cDNA was incorporatedinto the plant genome and was properly expressed inplants. Western blot analysis showed that the 80 kDa fulllength hLf protein was produced in transgenic tuberous roots. Overall results indicated that sweet potato would be anexcellent host to produce human therapeutic proteins.

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