Composite
Part:BBa_K3895007
Designed by: Rui Zhu Group: iGEM21_SZ_SHD (2021-10-01)
kerAvDZ50_dna_pET-28a(+)
Construction
Keratinase KerAVDZ50 (BBa_K3895003) is an extracellular serine thiol alkaline protease from the Actinomadura viridilutea strain, which can be isolated from the Algerian fishing port [1]. As a thermo-alkaline keratinases, it can function at a pH range of 7–12 and a temperature range of 35–80 °C. The keratinase was compared with the other 3 keratinases (BBa_K3895004; BBa_K3895005;BBa_K3895006) with the same construction method. 6x His-tags were added to both sides of the sequence for purification and constructed into PET-28a(+). The constitutive T7 promoter (BBa_I719005) derived from the T7 bacteriophage allows high expression of proteins.
Protein purification
Name of keratinase | IPTG concentration/mM | Temperature/°C | Time/h |
---|---|---|---|
KerAVDZ50 | 0.67139 | 16 | 12 |
Protein Concentration
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 606
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 411
Illegal AgeI site found at 1149 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 351
[edit]
Categories
Parameters
None |