Difference between revisions of "Part:BBa K2933179"

 
Line 18: Line 18:
 
<partinfo>BBa_K2933179 parameters</partinfo>
 
<partinfo>BBa_K2933179 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
===Usage and Biology===
 +
<br>
 +
 +
===Molecular cloning===
 +
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br>
 +
<p style="text-align: center;">
 +
  [[File:IMP-71-PCR.png|400px]]<br>
 +
'''Figure 1.'''  Left: The PCR result of IMP-71. Right: The verification results by enzyme digestion.<br>
 +
</p>
 +
After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br>

Revision as of 12:53, 21 September 2019


Tac promoter+RBS a+Linker g+GST+Linker e+IMP-71

This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+IMP-71),and the biological module can be built into E.coli for protein expression.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1518
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1518
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1518
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1518
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 181


Usage and Biology


Molecular cloning

First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

IMP-71-PCR.png
Figure 1. Left: The PCR result of IMP-71. Right: The verification results by enzyme digestion.

After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.