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Gal4-UAS System in Drosophila melanogaster
생화학실험
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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.
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Gal4-UAS System
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Gal4-UAS System ♀ ♂ Cg-Gal4 UAS-GFP Genomic enhancer GAL4 UAS Gene X
♀ ♂ Cg>GFP
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Gal4-UAS System Genomic enhancer GAL4 UAS Gene X Protein X Protein X
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Confocal Microscopy (공초점 현미경)
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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.
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Western blot sample preparation
Eppendorf tube에 lysis buffer를 넣는다. Larva를 selection해서 tube에 넣는다. Homogenization한다. -70℃에 보관한다.
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