Summer research

1
Summer research

• ????Yeast One Hybrid system
• ??????????? ?????

??04? 881714 ???
2
Institute introduction

• ?????
• ??????
• ????????
• ??????

3
Introduction
B cell (Bone marrow)

T cell(Thymus)
Lymphocyte stem cell (Bone marrow)
Tc (Cytotxic T cell)
Th (Helper T cell)
4
(No Transcript)
5
Introduction

• Th1 cells activate macrophages. Enabling them to
  destroy intracellular microorganisms more
  efficiently.
• Th2 cells drive B cells to differentiate and
  produce immunoglobulins of all other type.

6
(No Transcript)
7
Introduction

• T????????Th0????? Th1????????,?Interferon gamma
  promoter (pIFN-?) ???????IFN-?????
• T-bet ???? T??? Th0 ????? Th1 ??,????? IFN-? ????

8
Introduction

9
Material

• Yeast

10
Main difference between yeast and E.coli culture

• Incubation time
• Incubation temperature
• Culture medium

Yeast E.Coli
46 days 16 hours
Yeast E.Coli
30? 37?
Yeast E.Coli
YPD LB
11
Method

• Yeast One Hybrid system

12

• One-Hybrid System provides an in vivo genetic
  assay used for isolating novel genes encoding
  proteins that bind to a target, cis-acting
  regulatory element
• Protein-DNA interaction

13

• ?? T-bet ?? IFN-???????????,????? T-bet ?
  IFN-?????????? Yeast one hybrid
  ??????????????????,??????????????????
  IFN-?????????,???????????????? T cell ? Th0 ?????
  Th1 ???

14
(No Transcript)
15
(No Transcript)
16
(No Transcript)
17
Yeast strainYM4271(MATa)?Y187(MATa) Cant
synthesis histidine? leucine?uracil by itself?
Incubate
Yeast selection medium -His or -Leu or -Ura
Yeast
Yeast cant growth on selection medium
18
(No Transcript)
19
3-ATa inhibitor of the HIS3 gene product
Appropriate 3-AT
Yeast selection medium -His
20
Incubate
Yeast selection medium -His/-Leu
Yeast cant growth
Incubate
Yeast selection medium -His/-Leu
21
Appropriate 3-AT
Yeast selection medium -His/-Leu
22
Yeast mating
Yeast selection medium -His/-Leu
23
YM4271
Y187
24
Future work

• Screen library to find new transcription factor.

25
THE END Thank you