Gateway Cloning System

소개글

data club에서 발표한 Gateway Cloning System에 대한 발표자료입니다.

목차

- Cloning

- Vector

- Origin & Promoter

- Promoter

- The Cloning Bottleneck

- Designing a New Approach to Cloning

- Entry Vector System

- Applications

- LR & BP reaction

- Manufacturing process of Entry clone

- LR reaction

- Destination vector

- N-term tag

- Tagging

- N or C-term Fusion

- Tag on demand system

본문내용

1. Plasmid vector
Self replication
Circular DNA
Double strand
2. Virus vector
Bacteriophage
Origin & Promoter
Origin
pUC, f1, … origin
High or low copy number replication
Promoter
Recognize the RNA polymerase
Expression vector
E.coli : lac, trp, tac, λPL
Mammalian : SV40,CMV, EF1α, phosphoglycerate kinase,…
Promoter
In EPC(Endothelial Progenitor Cell)
CMV(Cytomegalovirus) in Lentivirus
EF1α(Elongation factor 1α) in Lentivirus
PGK(Phosphoglycerate kinase) in Lentivirus
CMV>EF1α>PGK≒0
In Mouse Embryonic stem cell
EF1α in pRC
CMV in pcDNA3
RSV (Rous sarcoma virus) in pRC
EF1α>CMV>RSV≒0
STEM CELLs 2006;24:199-208
STEM CELLs 2003;21:647-653
The Cloning Bottleneck
5`

참고 자료

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