Difference between revisions of "Part:BBa K414005"
m |
|||
| Line 1: | Line 1: | ||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K414005 short</partinfo> | <partinfo>BBa_K414005 short</partinfo> | ||
| + | |||
| + | |||
| + | '''''<center><span style="font-weight:bold; font-size:110%; color:red">Due to patent issues, samples of this part cannot be requested. - iGEM Headquarters</span></center>''''' | ||
| + | |||
The ccdB gene is a topoisomerase II poison and will kill most cell lines. This is an improvement to the prior ccdB cell death gene (BBa_P1010) in that the size has been reduced and it is now easier to use. CcdB can be used as a selectable marker by mixing it with the desired structure and a plasmid cut with BioBrick restriction enzymes. After ligation and transformation, because ccdB is toxic to most cell lines, the only visible colonies should contain the desired structure in the desired plasmid. CcdB is toxic to most cell lines with the exception of DB3.1 cell lines (BBa_V1005). The part has two point mutations from the original given sequence of the pK7FWG2, however it has been shown that these mutations do not affect translation of ccdB. | The ccdB gene is a topoisomerase II poison and will kill most cell lines. This is an improvement to the prior ccdB cell death gene (BBa_P1010) in that the size has been reduced and it is now easier to use. CcdB can be used as a selectable marker by mixing it with the desired structure and a plasmid cut with BioBrick restriction enzymes. After ligation and transformation, because ccdB is toxic to most cell lines, the only visible colonies should contain the desired structure in the desired plasmid. CcdB is toxic to most cell lines with the exception of DB3.1 cell lines (BBa_V1005). The part has two point mutations from the original given sequence of the pK7FWG2, however it has been shown that these mutations do not affect translation of ccdB. | ||
Revision as of 20:31, 3 July 2014
ccdB minimal gene
The ccdB gene is a topoisomerase II poison and will kill most cell lines. This is an improvement to the prior ccdB cell death gene (BBa_P1010) in that the size has been reduced and it is now easier to use. CcdB can be used as a selectable marker by mixing it with the desired structure and a plasmid cut with BioBrick restriction enzymes. After ligation and transformation, because ccdB is toxic to most cell lines, the only visible colonies should contain the desired structure in the desired plasmid. CcdB is toxic to most cell lines with the exception of DB3.1 cell lines (BBa_V1005). The part has two point mutations from the original given sequence of the pK7FWG2, however it has been shown that these mutations do not affect translation of ccdB.
Cell Transformation plates indicating that two cell lines, NEB10 and Omnimax cell lines, are susceptible to the toxic effects of ccdB but still will grow in a cell line, DB3.1 cell lines, that are not effected by ccdB (left). Polymerase Chain Reaction gel indicating the successful amplification of ccdB (right).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 545