Part:BBa_K1922007

hsp70-p53

Introduction

Hsp70 promoter works after heat shock, in mouse, after the injection of gold nano rods, we use laser to realize the heat shock. Theoretically, increased temperature will lead the expression of luciferase, thus, we can get a feedback by injecting luciferase substrate.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 269
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 414
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 219
  Illegal BsaI.rc site found at 1321
  Illegal BsaI.rc site found at 2512
  Illegal SapI site found at 403
  

Construction

We do the restriction digest to make sure the plasmid has been constructed. ""

Function Test

After heat shock, the p53 linked with hsp was overexpressed, it means the promoter works. We hope that after heat shock, the hsp promoter can drive the transcription of the downstream p53. Transfected cells were treated with heat shock at 42 celsius degree for one hour, after one hour’s restoration in 37 celsius degree, protein was extracted for western blot. Our results show after heat shock, the expression level of p53 increased. ""

Hsp70-p53 was transfected cells, then we measured the cellular viability to prove that hsp70-p53 can help decreased the cellular viability in thermotherapy. ""