Gal4-UAS System in Drosophila melanogaster

Presentation on theme: "Gal4-UAS System in Drosophila melanogaster"— Presentation transcript:

1 Gal4-UAS System in Drosophila melanogaster
생화학실험

2 Gal4-UAS System The GAL4-UAS system is a biochemical method used to study gene expression and function in organisms such as the fruit fly. It was developed by Andrea Brand and Norbert Perrimon in 1993 and is considered a powerful technique for studying the expression of genes. The system has two parts: the GAL4 gene, encoding the yeast transcription activator protein GAL4, and the UAS (Upstream Activation Sequence), an enhancer to which GAL4 specifically binds to activate gene transcription.

3 Gal4-UAS System

4 Gal4-UAS System Genomic enhancer GAL4 UAS Gene X ♀ ♂

5 Gal4-UAS System Genomic enhancer GAL4 UAS Gene X Protein X Protein X

6 Confocal Microscopy (공초점 현미경)

7 Method (GAL4-UAS System )
1. Gal4 Virgin female을 harvest한다. 2. UAS-GFP male 과 교배시킨다. 3. Egg를 6시간 동안 받는다. (여기까진 각반 반장과 조교들이 이미진행) 4. 96h(A.E.L.)~120h(A.E.L) 정도 지난 larva를 Selection 한다. 5. 슬라이드 글라스에 larva를 올린다. 6. 형광 현미경으로 관찰한다.

8 Mini-prep확인 EcoRI GFP KpnI XbaI pUAST-GFP

9 Method ( Mini-prep 확인 ) 1. 2. 37℃, 20min 3. 전기영동으로 insert 확인
Plasmid-DNA 100ng Enzyme ( EcoRI / XbaI ) 0.5ul / 0.5ul 10x Reaction buffer 2ul T.D.W Up to 20ul Total 20ul