Difference between revisions of "Part:BBa K2933217"

Revision as of 14:34, 21 September 2019

T7 promoter+RBS b+Linker h+His+Linker f+VIM-66+T7 terminator

This part consists of T7 promoter, RBS and protein coding sequence（His+Linker f+VIM-66），and the biological module can be built into E.coli for protein expression.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 47
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 169
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 942
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 47
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 47
1000
COMPATIBLE WITH RFC[1000]

Usage and Biology=

This composite part is made up with three basic parts, the His tag, T7 promoter, RBS and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of VIM-66 . It is convenient for us to purify our target protein.

Molecular cloning

We insert VIM-66 gene into the standard vector then transfer it into E.coli.

Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification

@@ Line 22: / Line 22: @@
 ===Molecular cloning===
 We insert VIM-66 gene into the standard vector then transfer it into E.coli.
-    [[File:VIM-66-PCR.jpeg|600px|center|]]
+    [[File:VIM-66-PCR1.jpeg|600px|center|]]
 <p style="text-align: center;">
 '''Figure 1.''' Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification