Difference between revisions of "Part:BBa K3712013"
Line 15: | Line 15: | ||
</head> | </head> | ||
<body> | <body> | ||
− | < | + | <center><img src="https://2021.igem.org/wiki/images/4/45/T--HUST2-China--The_agarose_gel_electrophoresis_map.png" style="width:318px;height:472px"></center> |
<p><b>Figure 1</b> The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system. About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment. </p> | <p><b>Figure 1</b> The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system. About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment. </p> | ||
<br> | <br> |
Revision as of 08:52, 21 October 2021
pR/pL promotor
USAGE AND RESULTS
PR/PL, a tandem PR and PL promoter from lambda phage, to ensure high-level expression of genes.Our target gene was cloned downstream of the pL and/or pR promoters. When cultured at 37℃, the expression of target gene is inhibited by Tcl and the target protein is hardly expressed. Under the induction of temperature at 45℃ or infrared light which increases temperature above 37℃, Tcl degrades and the inhibition of the promoter releases. So, the expression of target protein is initiated
RESULTS
Figure 1 The agarose gel electrophoresis map of Wild-type BLP-7-27 ELP, Optimized BLP-7-27 ELP, Wild-type TLR2-27ELP, and Optimized TLR2-27 ELP respectively constructed in pR/pL expression system. About 200-1000 ng of plasmid was digested at 37℃ for 30-60 minutes and analyzed on 1% Agarose Gel. Lane1: circular plasmid (brighter) and supercoiled structure; Lane2: plasmid (brighter) digested by EcoRI and BamHI and target gene fragment.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 116
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]