Difference between revisions of "Part:BBa K578005"
m (→Use in Competent Cells) |
|||
(One intermediate revision by the same user not shown) | |||
Line 13: | Line 13: | ||
===Confirmation of Function=== | ===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. | |
[[Image: ASU_GFP_Duet.png|400px]] | [[Image: ASU_GFP_Duet.png|400px]] | ||
− | === | + | ===Expression in E.coli=== |
− | E. | + | 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. |
<!-- Uncomment this to enable Functional Parameter display | <!-- Uncomment this to enable Functional Parameter display |
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.