Difference between revisions of "Part:BBa K3895009"

Revision as of 13:45, 15 October 2021

kerBlMKU3_dna_pET-28a(+)

ggggg

Construction

Keratinase KerBlMKU3 (BBa_K3895005) is a serine keratinase originally secreted by Bacillus licheniformis MKU3. It can work at 37°C for 48 h in nutrient broth or on feather medium [1]. 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.

Name of keratinase	IPTG concentration/mM	Temperature/°C	Time/h
KerBIMKU3	0.4	37	4

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 606
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 411
Illegal AgeI site found at 1149
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 351

@@ Line 5: / Line 5: @@
 ggggg
+==Construction==
+Keratinase KerBlMKU3 (<partinfo>K3895005</partinfo>) is a serine keratinase originally secreted by ''Bacillus licheniformis MKU3''. It can work at 37°C for 48 h in nutrient broth or on feather medium [1]. The keratinase was compared with the other 3 keratinases (<partinfo>K3895004</partinfo>; <partinfo>K3895005</partinfo>;<partinfo>K3895006</partinfo>) 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 (<partinfo>I719005</partinfo>) derived from the T7 bacteriophage allows high expression of proteins.
+[[File:T--SZ SHD--KU3plasmid.jpg|400px|center]]
 {|border=1 width="90%" align="center"