Difference between revisions of "Part:BBa K3895007"

Revision as of 07:55, 15 October 2021

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. 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

Figure 2. PCR and gel electrophoresis of KerAVDZ50.

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 21: / Line 21: @@
 |}
-[[File:T--SZ_SHD--verify.png]]
 {|border=0 width="90%" align="center"
 |-align="center"
-|[[File:T--SZ SHD--Z50plasmid.jpg]]
+|[[File:T--SZ_SHD--verify.png]]
 |[[File:T--SZ SHD--PCR2.png|300px|thumb|center|''' Figure 2.''' PCR and gel electrophoresis of KerAVDZ50.]]
 |}