Difference between revisions of "Part:BBa K4166005"

Revision as of 02:57, 12 October 2022

Camp +8 mutation.

Runway plasmid replication

Plasmid Copy Number and Expression - Alma 2022

We created a series of mutants of BBa_J04450 in the origin of replication, and wanted to both quantify the copy number and see the effect the copy number has on gene expression.

We isolated both genomic DNA and plasmid DNA from the cells and ran a serial dilution of these on a gel to observe the following.

From the intensity of these bands we estimate the copy number of this origin at ~387 copies/cell. We found that this plasmid both increases copy number and expression.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
23
INCOMPATIBLE WITH RFC[23]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
25
INCOMPATIBLE WITH RFC[25]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.

@@ Line 4: / Line 4: @@
 Runway plasmid replication
+== Plasmid Copy Number and Expression - Alma 2022 ==
+We created a series of mutants of BBa_J04450 in the origin of replication, and wanted to both quantify the copy number and see the effect the copy number has on gene expression.
+We isolated both genomic DNA and plasmid DNA from the cells and ran a serial dilution of these on a gel to observe the following.
+[[File:T--Alma--gel_005.png|350px|right]]
+[[File:T--Alma--pcnPlate.png|350px|left]]
+From the intensity of these bands we estimate the copy number of this origin at ~387 copies/cell.
+We found that this plasmid both increases copy number and expression.
 <!-- Add more about the biology of this part here