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2018 Vol.36, Issue 3 Preview Page

Research Article

Efficiency of Vegetable Soft Rot Pathogen Pectobacterium carotovorum subsp. carotovorum(Pcc21) as a Host Cell for Foreign Gene Expression

채소무름병균 Pectobacterium carotovorum subsp. carotovorum(Pcc21)의 외래유전자 발현용 숙주세포로서의 효율성 비교

Won-Woo Bae1
,
Yong-Sun Moon1*
배 원우1
,
문 용선1*
1Department of Horticulture and Life Science, Yeungnam University
1영남대학교 원예생명과학과
*교신저자. *Corresponding Author. 
June 2018. pp. 426-434
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Abstract

Enhanced introduction of foreign genes into plants has been achieved through the development of expression vectors and the use of diverse host cells. Escherichia cili and Agrobacterium tumefaciens are commonly used for plant transformation. However, there is still high demand for development of new host systems for foreign protein production in biotechnology and medical industry. Recently Pectobacterium carotovorum subsp. carotovorum (Pcc21), a pathogen causing soft  rot disease in potato, has received attention for biotechnological use because of its ease to grow and transform. In this study, we examined the potential of P. carotovorum (Pcc21) as a novel host system by assessing the efficiency of GFP protein production compared with other common host systems E. coli (DH5α) and A. tume-faciens (LBA4404). GFP production was evaluated by both RT-PCR and immunoblotting. Pcc21 produced about 30% more GFP protein than did DH5α and LBA4404. We suggest that Pcc21 is a potent candidate host to be used for foreign protein production as well as to be used as substitute for A. tumefaciens for transient and/or stable transformation of plants due to its high efficiency in in-fecting plants.

Keywords
competent cells
gram negative bacteria
overexpression
soft rot disease
transient expression

외래유전자 형질발현을 증폭하기 위해 형질발현 벡터의 개발뿐만 아니라 박테리아, 곰팡이, 효모, 곤충, 식물 및 동물세포와 같은 다양한 숙주세포의 개발이 시도되어 왔다. 특히 DH5α, XL1-blue, Top10 및 BL21과 같은 대장균(Escherichia coli)는 단백질 생산과 형질전환에 가장 널리 사용되고 있는 숙주 중 하나이며, LBA4404, GV3101 및 EHA105와 같은 아그로박테리움(Agrobacterium tumefaciens)은 식물 형질전환에 흔히 사용되고 있다. 그러나 여전히 의학 및 생명공학 분야에서는 단백질 생산을 위한 새로운 숙주세포 개발에 대한 요구가 있다. 최근 감자 무름병균인 펙토박테리아(Pectobacterium carotovorum subsp. carotovorum)는 사용의 편리함 때문에 새로운 숙주세포로써의 가능성에 대한 관심을 얻게 되었다. 이번 연구에서 E. coli(DH5α), A. tumefaciens(LBA4404) 및 P. carotovorum(Pcc21)을 포함하는 다양한 숙주세포에서 GFP 단백질생산 효율을 조사하였다. GFP 전사량은 RT-PCR로 확인하였고 단백질은 Western blot 분석으로 분석하였다. 그 결과 Pcc21은 DH5α와 LBA 4404와 비교했을 때 30% 정도 더 높은 GFP 단백질을 생산하였고 전사량 또한 증가하였다. 따라서 펙토박테리아 Pcc21이 외래 단백질 발현을 위한 새로운 숙주세포로써의 가능성이 높은 것으로 생각된다. 더 나아가 펙토박테리아는 쉽게 식물에 감염되기 때문에 A. tumefaciens 대신 식물의 일시적, 영구적 형질전환을 위한 숙주세포로 사용할 수 있을 것이라 생각된다.

키워드
컴피턴트 세포
그램 음성균
과 발현
무름병
일시적 발현
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Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :원예과학기술지
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 36
  • No :3
  • Pages :426-434
  • Received Date : 2017-06-28
  • Revised Date : 2017-09-04
  • Accepted Date : 2017-09-22
  • DOI :https://doi.org/10.12972/kjhst.20180042
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