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 projectMy project

KO 老鼠的建立Lupz 基因的研究老鼠的行為

KOKO 老鼠老鼠 nn promoterrtTA and TRE systemCre/loxP system

nn promoternn promoter

nn:

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

rtTA & TRE systemrtTA & TRE system

Cre/loxP systemCre/loxP system

Cre/loxP is the recombinase system of bacteriophage p1.

Cre recognizes and catalyzes recombination between 34bps loxP site.

Cre recombinationCre recombination

Combine allCombine all

nn rtTA Cre TRE

nn : tissue-specificrtTA & TRE : doxycycline induceCre/loxP : turn on knock-out

How to apply on mice?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?What is the advantage?

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

nnC4 rtTA

TRECre ScaIScaIH

M

9-1

9-2

9-3

9-4

9-5

10-2

10-3

10-4

10-5

11-3

11-4*

LM

**

HM

9-1

9-2

9-3

9-4

9-5

10-2

10-3

10-4

10-5

11-3

11-4*

LM

**

EcoRI EcoRI EcoRI

1.9kb

6.0kb

1.9kb 3.8kb

3.8kb

LuzpLuzp

Luzp encodes a protein (LUZP) expressing in brain areas such as hippocampus and cerebellum .

Subcellular localization is also revealed by immunocytochemistry(ICC) .

ICC localizationICC localization

What’s wrong with What’s wrong with LuzpLuzp KO mice? KO mice? - Neural tube closure defect - Neural tube closure defect

-/- +/- +/+

-/- +/- +/+ +/- -/- +/+

+/+ +/- +/- -/- +/-

Exencephaly in Exencephaly in LuzpLuzp-/- mice -/- mice through E10.5 to E14.5 through E10.5 to E14.5

E10.5 E11.5E14.5

Behavior effectBehavior effect

What did I do in summer? What did I do in summer?

Southern blotVector construct PCRICCFeed mice and clean their room ^^|||