Constructing the Plasmids: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[IGEM:MIT/2009/OfficialWiki]] | [[IGEM:MIT/2009/OfficialWiki]] | ||
Construction of the PhyB-CFP and Pif3-YFP Plasmids began with the amplification of each individual | Construction of the PhyB-CFP and Pif3-YFP Plasmids began with the amplification of each individual sequences, PhyB, CFP, Pif3 and YFP, from the Berkley Lab Plasmids. Through Fusion PCR the PhyB-CFP and Pif3-YFP sequences were then created. These sequence were subsequently inserted into the prS313 backbone, a plasmid which contains a constitutive Myo2 promoter and the His gene. | ||
A variety of C and N terminus localization Sequences were chosen to test the PhyB, Pif System. Each localization sequence, once amplified, was cloned into the appropriate backbone. N terminus tags were destined for the Pif3-YPF backbone and C terminus for the PhyB-CFP Backbone. | |||
Revision as of 08:09, 16 July 2009
Construction of the PhyB-CFP and Pif3-YFP Plasmids began with the amplification of each individual sequences, PhyB, CFP, Pif3 and YFP, from the Berkley Lab Plasmids. Through Fusion PCR the PhyB-CFP and Pif3-YFP sequences were then created. These sequence were subsequently inserted into the prS313 backbone, a plasmid which contains a constitutive Myo2 promoter and the His gene.
A variety of C and N terminus localization Sequences were chosen to test the PhyB, Pif System. Each localization sequence, once amplified, was cloned into the appropriate backbone. N terminus tags were destined for the Pif3-YPF backbone and C terminus for the PhyB-CFP Backbone.
