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 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 Cre/loxP system

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

rtTA & TRE system

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

Cre recombination

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

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.

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

nnC4rtTA TRECre ScaI HM * LM ** HM * LM ** EcoRI 1.9kb 6.0kb 1.9kb3.8kb

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

ICC localization

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

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

Behavior effect

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