표제지
목차
I. 서론 7
II. 재료 및 방법 13
1. 재조합 운반체의 제작 및 식물체 재분화 13
2. Genomic DNA 분리 18
3. Polymerase Chain Reaction (PCR) 19
4. Reverse transcriptase (RT)-PCR 20
5. Genomic southern blot hybridization 20
6. Northern blot hybridization 21
7. 메티오닌 함량 분석 22
III. 결과 및 고찰 23
1. PrLeg 유전자가 도입된 형질전환 감자의 선발 23
2. 형질전환체의 분자생물학적 확인 24
3. Genomic southern blot hybridization을 통한 형질전환된 감자의 게놈 내에 삽입된 PrLeg cDNA의 확인 29
4. 형질전환된 감자 괴경에서의 메티오닌 함량 분석 32
IV. 결론 37
참고문헌 39
Abstract 49
감사의 글 51
Table 1. Composition of growth medium II 17
Table 2. Primer list for PCR of RT-PCR 19
Table 3. Selection efficiency of transformants by PPT 23
Fig. 1. The schematic diagram of recombinant T-DNA fragments bearing PrLeg cDNA in binary vector. 14
Fig. 2. PCR amplification result with transgenic potato. Each number indicates an individual transgenic plant. M, size marker; P, positive control; N, negative control, wild type Jowon. 25
Fig. 3. RT-PCR result with total RNA from candidate transformants. 26
Fig. 4. Northern blot hybridization for the identification of transcript of Bar or PrLeg gene in transgenic potato. Each number represents a transgenic plant. JW, Jowon (wild type). 28
Fig. 5. Southern blot hybridization of candidate transformants with CaMV35S::PrLeg recombinant vector. 30
Fig. 6. Methionine content in total protein fraction of transgenic potato tubers. 33