Difference between revisions of "Part:BBa K578005"

Latest revision as of 22:19, 5 October 2011

J23102+E0840 (Constitutive GFP)

This is a combination of the GFP coding region (E0840), and the medium strength constitutive promoter, J23102.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 708

Confirmation of Function

BBa_K578005 was successfully subcloned into a variety of vectors, including pSB1C3, pSB1A3, pSB1K3, and pRSFDuet-1 (Novagen). The green fluorescent phenotype was observed. The image below show transforments of BL21(DE3) on the right and K12 MG1655 on the left, both with BBa_K578005 in pRSFDuet-1. Both show visible constitutive expression of GFP.

Expression in E.coli

These results indicate BBa_K578005 shows a high level of constitutive expression in both E.coli K12 MG1655 and E.coli BL21(DE3), allowing it to function as a visible marker for linked genetic elements, or as a visual screening tool for vector religation.

@@ Line 17: / Line 17: @@
 [[Image: ASU_GFP_Duet.png|400px]]
-===Use in Competent Cells===
+===Expression in E.coli===
-E. Coli MG1655 with this BioBrick in pRSF Duet was successfully made chemically competent using the OWW TSS procedure, allowing for the transformation of a second plasmid.
+These results indicate BBa_K578005 shows a high level of constitutive expression in both E.coli K12 MG1655 and E.coli BL21(DE3), allowing it to function as a visible marker for linked genetic elements, or as a visual screening tool for vector religation.
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