Difference between revisions of "Part:BBa K4509420"
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<partinfo>BBa_K4509420 short</partinfo> | <partinfo>BBa_K4509420 short</partinfo> | ||
− | The enzyme horseradish peroxidase, found in the roots of horseradish catalyzes the oxidation of various organic substrates by hydrogen peroxide. It is a metalloenzyme with many isoforms | + | The enzyme horseradish peroxidase, found in the roots of horseradish catalyzes the oxidation of various organic substrates by hydrogen peroxide. It is a metalloenzyme with many isoforms; the most studied type is C. The composite part starts with the J23116 constitutive promoter, continues with RBS and is followed by the Horseradish Peroxidase coding sequence. The sequence ends with a terminator and a BamH1 restriction site. |
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==Characterization== | ==Characterization== | ||
+ | |||
+ | The promoter J23119 was replaced with J23116 and transformed into E.coli K12 MG1655. The Transformed colonies were cultured and OD was taken every 30 mins to plot the growth curve of the cells. | ||
+ | |||
+ | https://static.igem.wiki/teams/4509/wiki/registry/hrp-116-1.png | ||
+ | |||
+ | The growth curve for the HRP plasmid with J23119, J23116, J23106 and J23100 is plotted in this graph. The burden on cells are indirectly proportional to cell viability. Looking at the growth curve it may be observed that the growth of J23116 is much efficient than J23119. Thus the part has an improved cell growth | ||
+ | |||
+ | ==Cell Burden== | ||
+ | |||
+ | J23116 has a burden range of -4.5 ± 2.8% |
Revision as of 03:41, 12 October 2022
HORSERADISH PEROXIDASE with constitutive promoter J23116
The enzyme horseradish peroxidase, found in the roots of horseradish catalyzes the oxidation of various organic substrates by hydrogen peroxide. It is a metalloenzyme with many isoforms; the most studied type is C. The composite part starts with the J23116 constitutive promoter, continues with RBS and is followed by the Horseradish Peroxidase coding sequence. The sequence ends with a terminator and a BamH1 restriction site.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 445
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 565
Illegal AgeI site found at 719
Illegal AgeI site found at 1022 - 1000COMPATIBLE WITH RFC[1000]
Characterization
The promoter J23119 was replaced with J23116 and transformed into E.coli K12 MG1655. The Transformed colonies were cultured and OD was taken every 30 mins to plot the growth curve of the cells.
The growth curve for the HRP plasmid with J23119, J23116, J23106 and J23100 is plotted in this graph. The burden on cells are indirectly proportional to cell viability. Looking at the growth curve it may be observed that the growth of J23116 is much efficient than J23119. Thus the part has an improved cell growth
Cell Burden
J23116 has a burden range of -4.5 ± 2.8%