Difference between revisions of "Part:BBa K2933158"

Revision as of 08:04, 22 September 2019

His+Linker f+BlaB-14

This part encodes the fusion protein of His tag and BlaB-14 to promote the expression and purification of target protein(BlaB-14).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 51
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 647
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Usage and Biology

This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein BlaB-14. It encodes a protein which is BlaB-14 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.

Molecular cloning

First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.We then extracted plasmids and performed double enzyme digestion verification.

Figure 1. The PCR result of BlaB-14.

@@ Line 17: / Line 17: @@
 <partinfo>BBa_K2933158 parameters</partinfo>
 <!-- -->
+===Usage and Biology===
+This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein BlaB-14. It encodes a protein which is BlaB-14 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.
+===Molecular cloning===
+First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.We then extracted plasmids and performed double enzyme digestion verification.<br>
+<p style="text-align: center;">
+   [[File:Blab PCR.png|200px]]<br>
+'''Figure 1.'''  The PCR result of BlaB-14.<br>
+</p>