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My summer life in Yang-Ming

My summer life in Yang-Ming. 微生物及免疫學研究所 張南驥老師 神經科學研究所 錢嘉韻老師. 生科04級 891632 曾煥昌. 實驗室簡介. 張南驥老師 1.巨噬細胞分泌性蛋白 ( YM-1). 2.神經元 LUZP 蛋白. 錢嘉韻老師 1.α2 腎上腺素受體位置及功能研究. 2. 建立 TG & KO mice 來做基因標的. My project. KO 老鼠的建立 Lupz 基因的研究 老鼠的行為. KO 老鼠. nn promoter rtTA and TRE system

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My summer life in Yang-Ming

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  1. My summer life in Yang-Ming 微生物及免疫學研究所 張南驥老師 神經科學研究所 錢嘉韻老師 生科04級 891632 曾煥昌

  2. 實驗室簡介 張南驥老師 1.巨噬細胞分泌性蛋白 (YM-1). 2.神經元 LUZP 蛋白. 錢嘉韻老師 1.α2 腎上腺素受體位置及功能研究. 2.建立 TG & KO mice 來做基因標的.

  3. My project • KO 老鼠的建立 • Lupz 基因的研究 • 老鼠的行為

  4. KO老鼠 • nn promoter • rtTA and TRE system • Cre/loxP system

  5. nn promoter • nn: the promoter of α2C4 , one kind of the recepter of epinephrine.

  6. rtTA & TRE system

  7. Cre/loxP system • Cre/loxP is the recombinase system of bacteriophage p1. • Cre recognizes and catalyzes recombination between 34bps loxP site.

  8. Cre recombination

  9. Combine all nn rtTA Cre TRE nn : tissue-specific rtTA & TRE : doxycycline induce Cre/loxP : turn on knock-out

  10. How to apply on mice? • Do microinjection on the mice, get transgenic mice(chimeric mice). • Mate the mice to get hetozygotes. • Mate hetrozygotes to get homozygotes.

  11. What is the advantage? The target gene may be lethal. The knock out mice can be survival until we turn on the system.

  12. EcoRI EcoRI EcoRI 1.9kb 3.8kb ScaI ScaI nnC4 rtTA Cre TRE * * * * * * LM LM HM 9-1 9-2 9-3 9-4 9-5 10-2 10-3 10-4 10-5 11-3 11-4 HM 9-1 9-2 9-3 9-4 9-5 10-2 10-3 10-4 10-5 11-3 11-4 6.0kb 3.8kb 1.9kb

  13. Luzp • Luzp encodes a protein (LUZP) expressingin brain areas such as hippocampus and cerebellum . • Subcellular localization is also revealed by immunocytochemistry(ICC) .

  14. ICC localization

  15. +/+ +/- +/- -/- +/- -/- +/- +/+ -/- +/- +/+ +/- -/- +/+ What’s wrong with Luzp KO mice? - Neural tube closure defect

  16. E10.5 E11.5 E14.5 Exencephaly in Luzp-/- mice through E10.5 to E14.5

  17. Behavior effect

  18. What did I do in summer? • Southern blot • Vector construct • PCR • ICC • Feed mice and clean their room ^^|||

  19. The End

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