Difference between revisions of "Part:BBa K118022"
Line 13: | Line 13: | ||
*'''Group:''' [http://2018.igem.org/Team:UESTC-China iGEM Team UESTC-China 2018] | *'''Group:''' [http://2018.igem.org/Team:UESTC-China iGEM Team UESTC-China 2018] | ||
*'''Author:''' Liang Zhao, Yetao Zou | *'''Author:''' Liang Zhao, Yetao Zou | ||
− | *'''Summary:''' | + | *'''Summary:''' Enzyme digestion and enzyme activity assay |
===Characterization from iGEM18-UESTC-China=== | ===Characterization from iGEM18-UESTC-China=== | ||
Line 20: | Line 20: | ||
====Enzyme digestion==== | ====Enzyme digestion==== | ||
[[File:T--UESTC-China--Enzyme digestion|500px|thumb|center|'''Fig. 1''' ]] | [[File:T--UESTC-China--Enzyme digestion|500px|thumb|center|'''Fig. 1''' ]] | ||
− | ==== | + | ==== ==== |
We constructed a plasmid containing cenA gene [https://parts.igem.org/Part:BBa_K118023 BBa_K118023], cex gene [https://parts.igem.org/Part:BBa_K118022 BBa_K118022]. We transformed this plasmid into BL21(DE3). | We constructed a plasmid containing cenA gene [https://parts.igem.org/Part:BBa_K118023 BBa_K118023], cex gene [https://parts.igem.org/Part:BBa_K118022 BBa_K118022]. We transformed this plasmid into BL21(DE3). | ||
We used an intracellular fraction (crude enzyme solution) obtained by ultrasonication to carry out an experiment for measuring total enzyme activities by the method of filter paper assay. [[File:T--UESTC-China—total cellulase activity|500px|thumb|center|'''Fig. 2''' Enzyme activity of the total cellulase at pH7.0, 40 ℃]] | We used an intracellular fraction (crude enzyme solution) obtained by ultrasonication to carry out an experiment for measuring total enzyme activities by the method of filter paper assay. [[File:T--UESTC-China—total cellulase activity|500px|thumb|center|'''Fig. 2''' Enzyme activity of the total cellulase at pH7.0, 40 ℃]] |
Revision as of 08:49, 17 October 2018
cex coding sequence encoding Cellulomonas fimi exoglucanase
The cellulolytic bacterium Cellulomonas fimi uses an exoglucanase (from cex, accession M15824) along with 3 endoglucanases in the degradation of cellulose into cellobiose, before use B-glucosidase to catalyse the conversion of cellobiose to D-glucose.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 524
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 157
Illegal NgoMIV site found at 530
Illegal NgoMIV site found at 1032 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 577
Illegal SapI.rc site found at 660
Contribution
- Group: [http://2018.igem.org/Team:UESTC-China iGEM Team UESTC-China 2018]
- Author: Liang Zhao, Yetao Zou
- Summary: Enzyme digestion and enzyme activity assay
Characterization from iGEM18-UESTC-China
Molecular weight
This gene codes for a protein of 485 amino acids with a molecular mass of 51.2 kDa.
Enzyme digestion
We constructed a plasmid containing cenA gene BBa_K118023, cex gene BBa_K118022. We transformed this plasmid into BL21(DE3).
We used an intracellular fraction (crude enzyme solution) obtained by ultrasonication to carry out an experiment for measuring total enzyme activities by the method of filter paper assay.