KR101773338B1 - 중쇄지방산 생산에 이용되는 신규 유전자 및 이의 용도 - Google Patents
중쇄지방산 생산에 이용되는 신규 유전자 및 이의 용도 Download PDFInfo
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- KR101773338B1 KR101773338B1 KR1020150129738A KR20150129738A KR101773338B1 KR 101773338 B1 KR101773338 B1 KR 101773338B1 KR 1020150129738 A KR1020150129738 A KR 1020150129738A KR 20150129738 A KR20150129738 A KR 20150129738A KR 101773338 B1 KR101773338 B1 KR 101773338B1
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Abstract
Description
도 2는 쿠페아(Cuphea paucipetala) 유래 중쇄지방산 생산 유전자의 염기 서열(서열번호 2) 및 아미노산 서열(서열번호 1)을 나타낸 도이다.
도 3은 중쇄지방산 생산 유전자와 다른 식물체로부터 분리된 FatB 유전자들과의 아미노산 유연관계를 나타낸 도이다; 가로축은 100개의 아미노산 당 서로 다른 아미노산의 개수를 의미.
도 4는 중쇄지방산 생산 유전자를 포함하는 식물 형질전환용 운반체의 모식도이다.
도 5는 중쇄지방산 생산 유전자로 형질전환된 애기장대 형질전환체 A7 계통의 지방산 조성별 분포도이다; 대조구: 애기장대 비형질전환체.
도 6은 중쇄지방산 생산 유전자로 형질전환된 애기장대 형질전환체 A7 계통의 중쇄지방산 및 장쇄지방산 분포도이다; 대조구: 애기장대 비형질전환체.
도 7은 중쇄지방산 생산 유전자로 형질전환된 애기장대 형질전환체 A7 계통의 GC 분석에 의한 지방산 조성 그래프를 나타낸 도이다; (위) 대조구, (아래) A10 계통. IS; Internal Standard.
계통번호 | 중쇄지방산 | 장쇄지방산 | 중쇄지방산 | 장쇄지방산 | ||||||||
C10:0 | C12:0 | C14:0 | C16:0 | C18:0 | C18:1 | C18:2 | C18:3 | C20:0 | C20:1 | |||
대조구 | 0 | 0 | 0 | 8 | 3 | 17 | 29 | 19 | 2 | 21 | 0 | 100 |
A1 | 3 | 2 | 1 | 14 | 4 | 17 | 25 | 20 | 2 | 11 | 6 | 94 |
A2 | 4 | 1 | 1 | 15 | 3 | 14 | 26 | 20 | 3 | 12 | 6 | 94 |
A3 | 2 | 1 | 1 | 12 | 3 | 17 | 28 | 19 | 2 | 16 | 4 | 96 |
A4 | 5 | 3 | 1 | 14 | 3 | 18 | 23 | 17 | 2 | 12 | 9 | 91 |
A5 | 2 | 1 | 1 | 11 | 3 | 18 | 28 | 18 | 2 | 17 | 4 | 96 |
A6 | 5 | 2 | 1 | 14 | 3 | 15 | 25 | 20 | 2 | 12 | 8 | 92 |
A7 | 6 | 2 | 1 | 14 | 3 | 17 | 24 | 18 | 3 | 12 | 9 | 91 |
A8 | 2 | 1 | 1 | 12 | 3 | 17 | 26 | 17 | 2 | 18 | 4 | 96 |
A9 | 4 | 1 | 1 | 12 | 3 | 18 | 25 | 19 | 1 | 15 | 6 | 94 |
A10 | 4 | 1 | 1 | 14 | 3 | 15 | 26 | 19 | 3 | 13 | 6 | 94 |
A11 | 2 | 1 | 1 | 12 | 3 | 17 | 28 | 19 | 2 | 16 | 4 | 96 |
A12 | 5 | 2 | 1 | 14 | 3 | 17 | 24 | 18 | 2 | 12 | 9 | 91 |
계통번호 | 중쇄지방산 | 장쇄지방산 | 중쇄지방산 | 장쇄지방산 | ||||||||
C10:0 | C12:0 | C14:0 | C16:0 | C18:0 | C18:1 | C18:2 | C18:3 | C20:0 | C20:1 | |||
대조구 | 0 | 0 | 0 | 5 | 2 | 66 | 19 | 6 | 1 | 1 | 0 | 100 |
B1 | 3 | 1 | 1 | 11 | 4 | 56 | 18 | 6 | 0 | 0 | 5 | 95 |
B2 | 4 | 1 | 1 | 9 | 3 | 57 | 17 | 5 | 1 | 1 | 6 | 94 |
B3 | 6 | 2 | 1 | 11 | 2 | 53 | 17 | 5 | 1 | 1 | 9 | 91 |
B4 | 3 | 1 | 1 | 10 | 3 | 58 | 16 | 4 | 1 | 1 | 5 | 95 |
B5 | 4 | 1 | 1 | 10 | 3 | 55 | 17 | 5 | 8 | 1 | 6 | 94 |
B6 | 4 | 1 | 0 | 10 | 4 | 57 | 15 | 5 | 2 | 1 | 5 | 95 |
Claims (13)
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- (1) 서열번호 3 및 4로 표시되는 프라이머 세트를 이용하여, 쿠페아(Cuphea paucipetala)의 미숙종자로부터 지방산 싸이오에스테라아제(Fatty acid thioesterase) 신규 유전자를 분리하는 단계;
(2) 상기 단계 (1)에서 분리된, 서열번호 2의 염기서열로 이루어지는 지방산 싸이오에스테라아제(Fatty acid thioesterase) 신규 유전자를 포함하는 재조합 벡터를 제조하는 단계;
(3) 상기 재조합 벡터를 아그로박테리움을 이용하여 애기장대 식물체 또는 유채 식물체에 형질전환하는 단계;
(4) 바스타 제초제를 살포하여 상기 지방산 싸이오에스테라아제(Fatty acid thioesterase) 신규 유전자가 발현된 애기장대 형질전환 식물체 또는 유채 형질전환 식물체를 선별하는 단계;
(5) 상기 선별된 형질전환 식물체로부터 T1 종자를 수득하는 단계; 및
(6) 상기 단계 (5)의 T1 종자 중에 카프산(Caprate, C10:0), 라우린산(Laurate, C12:0) 및 미리스틱산(Myristate, C14:0)의 함량이 증가한 계통을 선발하는 단계;를 포함하는, 종자 내 카프산(Caprate, C10:0), 라우린산(Laurate, C12:0) 및 미리스틱산(Myristate, C14:0) 함량이 증가된 애기장대 형질전환 식물체 또는 유채 형질전환 식물체로부터 카프산(Caprate, C10:0), 라우린산(Laurate, C12:0) 및 미리스틱산(Myristate, C14:0)을 생산하는 방법.
- 제11항에 있어서, 상기 지방산 싸이오에스테라아제(Fatty acid thioesterase) 신규 유전자는 서열번호 1의 아미노산 서열로 이루어진 단백질을 암호화하는 것인 방법.
- 제11항에 있어서, 상기 지방산 싸이오에스테라아제(Fatty acid thioesterase) 신규 유전자는 싸이오에스테라아제5-3(Thioesterase5-3)으로 명명된 것인 방법.
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