DNA 클로닝 (4 판)

최석용,노현주,이한길

대표작으로『DNA클로닝』이있다.

1클로닝이란?1
1.1클로닝의정의2
1.2새로운유전자의발굴3
1.3과거의클로닝5
1.4현재의클로닝6

2클로닝을위한준비9
2.1클로닝설계에유용한소프트웨어10
2.2벡터,플라스미드,컨스트럭트&코작염기서열(Kozakconsensussequence)13
2.3MCS(MultipleCloningSite)18
2.4제한효소(RestrictionEnzyme,RE)와스타활성(staractivity)22
2.5아가로즈젤전기영동을이용한DNA분석29
2.6LB(Luria-Bertani)플레이트만들기34
2.7CompetentCell37
2.8DNA의질량을mole수로바꾸는법41
2.9새로운플라스미드를받았을때42
2.10cDNA라이브러리44

3클로닝첫걸음47
3.1Cut&Paste48
3.2DNA염기서열분석&direct염기서열분석61
3.3클로닝을위한PCR&nestedPCR65
3.4Fill-in(Full&Partial)77
3.5상보적인점착말단(CompatibleCohesiveEnd)83
3.6메틸화(Methylation)85
3.7세절편연결(Three-pieceligation)93
3.8위치지정돌연변이(Site-directedmutagenesis)96
3.9TA클로닝&T-벡터의제작102
3.10TOPOTA클로닝108
3.11Gateway클로닝110
3.12식물형질전환(Planttransformation)벡터의구조114
3.13Rhizobiumradiobacter형질전환117

4클로닝다음걸음119
4.1유전자중간부분에다른DNA를삽입(insertion)하는방법120
4.2유전자중간부분을삭제(deletion)하는방법124
4.3유전자에에피톱택을삽입하는방법&CRISPR/Cas9126
4.4번역융합(TranslationalFusion)vs전사융합(TranscriptionalFusion)133

5클로닝으로인한두통을덜어주는방법135
5.1Goldengateassembly&Goldengate를이용해푸는직소퍼즐136
5.2In-FusionSLIC(In-FusionSequence-andLigation-IndependentCloning)142
5.3??T4DNAPolSLIC
(T4DNAPolymeraseSequence-andLigation-IndependentCloning)153
5.4RE-freeSLIC159
5.5EMP(ExponentialMegaprimingPCR)클로닝161
5.6MRF(ModifiedRestrictionFree)클로닝165
5.7Non-templatePCR클로닝168
5.8cDNA합성의뢰171

6클로닝마지막걸음173
6.1다른종에서유사한유전자(homolog)를클로닝하는방법174
6.2RACE(RapidAmplificationofcDNAEnds)178
6.3BAC재조합181
6.4OldTrick:부분절단185
6.5벡터의변경187
6.6클로닝이다됐다고생각했는데틀이동돌연변이가있을때191
6.7클로닝의실제:억세게운이없는경우193

7클로너에게주는조언205
7.1클로닝이안될때206
7.2클로닝의정리와기록209

심화학습을위한문헌213
약자215
찾아보기217