restriction enzyme과 ligase의 작용 메카니즘과 bacteria의 restriction modification system에 관한 완벽한 자료를 담고 있습니다.
1. DNA cloning methodology
2. restriction enzyme - general features
3. Three types of DNA ends generated by restriction enzymes
4. Converting a 5’/3’ overhang to a blunt end
5. DNA Ligase
6. Ligation of foreign DNA to plasmid vectors
7. How does the bacterium protect its own DNA from restriction cleavage?
8. Restriction Modification System
9. Restriction/Modification in E. coli
Restriction enzymes and DNA ligase play essential roles in DNA cloning. How is it that a bacterium that produces a restriction enzyme does not cut its own DNA? Describe some general features of restriction enzyme site. What are the three types of DNA ends that can be generated after cutting DNA with restriction enzymes? What reaction is catalyzed by DNA ligase?
DNA Cloning Methodology
Cut the cloning vector with R.E. of choice, eg Eco RI
Cut DNA of interest with same R.E. or R.E. yielding same sticky ends, e.g. Bam HI and Sau 3A
Mix the restrict