Difference between revisions of "Part:BBa K3196033"
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This part is an improvement of part Baa_K500002, which codes the Geneart Laccase. Geneart Laccase is a polyphenol oxidase, which belongs to the family of blue multicopper oxidases. Laccases oxidize a broad range of substrates, preferably phenolic compounds. Compering with part BBa_K500002, We add a productive signal peptide called FLO10-αpro before the gene.
 
This part is an improvement of part Baa_K500002, which codes the Geneart Laccase. Geneart Laccase is a polyphenol oxidase, which belongs to the family of blue multicopper oxidases. Laccases oxidize a broad range of substrates, preferably phenolic compounds. Compering with part BBa_K500002, We add a productive signal peptide called FLO10-αpro before the gene.
 
<h1>'''Experiments'''</h1>
 
<h1>'''Experiments'''</h1>
−
We insert the segment into pPIC9K plasmid.
+
We insert the segment into pPIC9K plasmid.
 +
[[File:T--HUST-China--2019-improvement construction.png|400px|thumb|center|Figure1:This is the construct process of the new part and the old part into pPIC9k plasmid. ]]
 
By using BamHⅠ and SnaBⅠ,we make a kick on the FLO10-pPIC9k and replace the FLO10 signal peptide with BBa_K500002 fragment(the fragment is amplified by PCR).
 
By using BamHⅠ and SnaBⅠ,we make a kick on the FLO10-pPIC9k and replace the FLO10 signal peptide with BBa_K500002 fragment(the fragment is amplified by PCR).
 
By using SnaBⅠand EcoRⅠ, we add BBa_K500002 fragment after FLO10 signal peptide.
 
By using SnaBⅠand EcoRⅠ, we add BBa_K500002 fragment after FLO10 signal peptide.
−
[[File:T--HUST-China--2019-improvement construction.png|400px|thumb|center|Figure1:This is the construct process of the new part and the old part into pPIC9k plasmid. ]]
+
 
  
 
Here are the DNA gel electrophoresis result:
 
Here are the DNA gel electrophoresis result:
−
[[File:T--HUST-China--2019-improvement construction.png|400px|thumb|center|Figure2:The construction is successful. ]]
+
[[File:T--HUST-China--2019-improvement-cloning PCR.jpg|400px|thumb|center|Figure2:The construction is successful. ]]

Revision as of 05:02, 20 October 2019


FLO10-αpro-Geneart Laccase

This part is an improvement of part Baa_K500002, which codes the Geneart Laccase. Geneart Laccase is a polyphenol oxidase, which belongs to the family of blue multicopper oxidases. Laccases oxidize a broad range of substrates, preferably phenolic compounds.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 349
    Illegal BglII site found at 886
    Illegal BglII site found at 1582
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

This part is an improvement of part Baa_K500002, which codes the Geneart Laccase. Geneart Laccase is a polyphenol oxidase, which belongs to the family of blue multicopper oxidases. Laccases oxidize a broad range of substrates, preferably phenolic compounds. Compering with part BBa_K500002, We add a productive signal peptide called FLO10-αpro before the gene.

Experiments

We insert the segment into pPIC9K plasmid.

Figure1:This is the construct process of the new part and the old part into pPIC9k plasmid.

By using BamHⅠ and SnaBⅠ,we make a kick on the FLO10-pPIC9k and replace the FLO10 signal peptide with BBa_K500002 fragment(the fragment is amplified by PCR). By using SnaBⅠand EcoRⅠ, we add BBa_K500002 fragment after FLO10 signal peptide.


Here are the DNA gel electrophoresis result:

Figure2:The construction is successful.