Difference between revisions of "Part:BBa I13453:Experience"
(→User Reviews) |
(→User Reviews) |
||
| Line 1: | Line 1: | ||
| − | |||
__NOTOC__ | __NOTOC__ | ||
This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
| Line 34: | Line 33: | ||
|width='60%' valign='top'| | |width='60%' valign='top'| | ||
'''pBad I13453''' confers expression of tetracycline resistance when placed upstream of '''hixC-RBS-Tet...''' (<partinfo>BBa_J44010</partinfo>). The presence of '''hixC''' (26 bp) between '''pBad''' and '''RBS-Tet''' does not disrupt transcription. In the insulated vector <partinfo>pSB1A7</partinfo> we have observed that tetracycline resistance depends upon the presence and proper orientation of '''pBad''' relative to the coding region (Davidson and Missouri Western 2006). '''pBad-hixC-RBS-Tet''' is tetracycline resistant, whereas '''RBS-Tet''' and '''pBad-hixC-Tet<sub>rev</sub>-RBS<sub>rev</sub>''' are not. We have observed Tet expression in the absence of L-arabinose (the affects of L-arabinose were not tested in this system). In summary, '''pBad I13453''' (which includes the Cap and AraC protein binding domains and polymerase binding site; Lee, PNAS, 1981) appears to be a functional constitutive promoter that does not depend upon L-arabinose for expression. | '''pBad I13453''' confers expression of tetracycline resistance when placed upstream of '''hixC-RBS-Tet...''' (<partinfo>BBa_J44010</partinfo>). The presence of '''hixC''' (26 bp) between '''pBad''' and '''RBS-Tet''' does not disrupt transcription. In the insulated vector <partinfo>pSB1A7</partinfo> we have observed that tetracycline resistance depends upon the presence and proper orientation of '''pBad''' relative to the coding region (Davidson and Missouri Western 2006). '''pBad-hixC-RBS-Tet''' is tetracycline resistant, whereas '''RBS-Tet''' and '''pBad-hixC-Tet<sub>rev</sub>-RBS<sub>rev</sub>''' are not. We have observed Tet expression in the absence of L-arabinose (the affects of L-arabinose were not tested in this system). In summary, '''pBad I13453''' (which includes the Cap and AraC protein binding domains and polymerase binding site; Lee, PNAS, 1981) appears to be a functional constitutive promoter that does not depend upon L-arabinose for expression. | ||
| + | |} | ||
| + | {|width='80%' style='border:1px solid gray' | ||
| + | |- | ||
| + | |width='10%'| | ||
| + | <partinfo>BBa_I13453 AddReview 5</partinfo> | ||
| + | <I>British_Columbia iGEM 2009</I> | ||
| + | |width='60%' valign='top'| | ||
| + | We performed extensive testing on <partinfo>I13453</partinfo> as part of our creation of a family of variable-strength arabinose-inducible promoters. We found that induction of <partinfo>I13453</partinfo> with L-arabinose led to expression of a coupled fluorescent reporter; contrary to the above review, we did not observe any reporter expression in the absence of L-arabinose. <partinfo>I13453</partinfo> response to L-arabinose was specific and dependent on input concentration. Our detailed experimental measurements for <partinfo>I13453</partinfo> can be found [[Part:BBa_I13453:British_Columbia|here]] and a comparison of the entire pBAD promoter family can be found [https://parts.igem.org/PBAD_Promoter_Family here]. | ||
|} | |} | ||
<!-- DON'T DELETE --><partinfo>BBa_I13453 EndReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_I13453 EndReviews</partinfo> | ||
Revision as of 01:25, 22 October 2009
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_I13453
User Reviews
UNIQ3ae0b64156864050-partinfo-00000000-QINU
|
•••••
Antiquity |
This review comes from the old result system and indicates that this part worked in some test. |
|
•••••
kahaynes |
pBad I13453 confers expression of tetracycline resistance when placed upstream of hixC-RBS-Tet... (BBa_J44010). The presence of hixC (26 bp) between pBad and RBS-Tet does not disrupt transcription. In the insulated vector pSB1A7 we have observed that tetracycline resistance depends upon the presence and proper orientation of pBad relative to the coding region (Davidson and Missouri Western 2006). pBad-hixC-RBS-Tet is tetracycline resistant, whereas RBS-Tet and pBad-hixC-Tetrev-RBSrev are not. We have observed Tet expression in the absence of L-arabinose (the affects of L-arabinose were not tested in this system). In summary, pBad I13453 (which includes the Cap and AraC protein binding domains and polymerase binding site; Lee, PNAS, 1981) appears to be a functional constitutive promoter that does not depend upon L-arabinose for expression. |
|
•••••
British_Columbia iGEM 2009 |
We performed extensive testing on BBa_I13453 as part of our creation of a family of variable-strength arabinose-inducible promoters. We found that induction of BBa_I13453 with L-arabinose led to expression of a coupled fluorescent reporter; contrary to the above review, we did not observe any reporter expression in the absence of L-arabinose. BBa_I13453 response to L-arabinose was specific and dependent on input concentration. Our detailed experimental measurements for BBa_I13453 can be found here and a comparison of the entire pBAD promoter family can be found here. |
UNIQ3ae0b64156864050-partinfo-0000000A-QINU