Difference between revisions of "Part:BBa K211001:Experience"

(→‎Applications of BBa_K211001)
(→‎Applications of BBa_K211001)
Line 6: Line 6:
  
 
Engineering GPCR signalling pathways.
 
Engineering GPCR signalling pathways.
 +
 +
===HKUST2009 lab experience of BBa_K211002===
 +
 +
HKUST team in 2009 has characterized the [https://parts.igem.org/Part:BBa_K211001 BBa_K211001] in the experiment shown below. This odorant sensing GPCR has been proven to sense its ligand hexanal in the heterologous ''Saccharomyces cerevisiae'' system.
 +
 +
1.Protocol
 +
 +
FACS
 +
 +
1. Transformed yeast cells are induced with galactose first for 1 hour.
 +
 +
2. Yeasts after 1 hour, at OD600 = 0.8 are induced with diacetyl solution at 5mM.
 +
 +
3. Every 1.5 hour we need to collect samples for FACS analaysis,both control or experiment groups.
 +
 +
4. FACS standard method for sample preparation.
 +
 +
5. Gel pictual for results.
 +
 +
(2)Result
 +
 +
 +
After being induced with galactose, the RI7 receptor is induced with its ligand hexanal. From 0 to 4 hours, the cells sense the ligand and trigger the downstream mating response, leading to a cell cycle arrest at G1 phase. As a result, the DNA content is shifted towards the G1 non-replicated DNA content.
 +
[[Image:Experiment2.PNG]]
 +
 +
 +
For more detailed information, please refer to our [http://2009.igem.org/Team:HKUST/OdorantSensing team Wiki].
  
 
===User Reviews===
 
===User Reviews===

Revision as of 22:31, 21 October 2009

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K211001

Engineering GPCR signalling pathways.

HKUST2009 lab experience of BBa_K211002

HKUST team in 2009 has characterized the BBa_K211001 in the experiment shown below. This odorant sensing GPCR has been proven to sense its ligand hexanal in the heterologous Saccharomyces cerevisiae system.

1.Protocol

FACS

1. Transformed yeast cells are induced with galactose first for 1 hour.

2. Yeasts after 1 hour, at OD600 = 0.8 are induced with diacetyl solution at 5mM.

3. Every 1.5 hour we need to collect samples for FACS analaysis,both control or experiment groups.

4. FACS standard method for sample preparation.

5. Gel pictual for results.

(2)Result


After being induced with galactose, the RI7 receptor is induced with its ligand hexanal. From 0 to 4 hours, the cells sense the ligand and trigger the downstream mating response, leading to a cell cycle arrest at G1 phase. As a result, the DNA content is shifted towards the G1 non-replicated DNA content. Experiment2.PNG


For more detailed information, please refer to our [http://2009.igem.org/Team:HKUST/OdorantSensing team Wiki].

User Reviews

UNIQd65bfeb9418f13ef-partinfo-00000000-QINU UNIQd65bfeb9418f13ef-partinfo-00000001-QINU