Difference between revisions of "Part:BBa K3867001"
 
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[[File:K3867001-1.jpg|center]]
 
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This gene was cloned into the vector pSB1C3. PCR method was used to identify. Fig.1 shows the result of identification.  
 
This gene was cloned into the vector pSB1C3. PCR method was used to identify. Fig.1 shows the result of identification.  
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[[File:K3867001-2.jpg|center]]
 
[[File:K3867001-2.jpg|center]]
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Fig.1. The result of PGM and UGPase gene cloning.
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Fig.1. The result of ADG1 gene cloning.
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M: Marker; 1: PCR result of PGM and UGPase; 2: Digestion of pSB1C3 containing PGM and UGPase genes.
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M: Marker; 1: plasmid of pSB1C3-ADG1; 2: PCR result of ADG1 gene.
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When this gene was constructed to the expression vector pETDuet-1, ADG1 protein was expressed. His tag was used to purify ADG1 protein. The identification result was showed in Fig.2
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[[File:K3867001-3.jpg|center]]
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Fig.2. The expression of ADG1 with vector pETDuet-1-ADG1 in E. coli.
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M, Marker; 1, Purification of ADG1 after Induced by IPTG; 2, Induced by IPTG; 3, No induced by IPTG.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 06:02, 14 October 2021


ADG1 （ADP glucose pyrophosphorylase 1)

ADP glucose pyrophosphorylase, abbreviated as AGPase, containing 2 subunits, ADG1 and APL1. ADG1 encodes the small subunit and APL1 encodes the large subunit. The large subunit plays a regulatory role whereas the small subunit (ADG1) is the catalytic isoform responsible for AGPase activity. AGPase catalyzes Glucose-6-phosphate (Glu-6-P) converted to adenosine diphosphate glucose (ADPG), which is a key enzyme in amylose synthesis and catalyzes the first, rate limiting step in amylose biosynthesis.

K3867001-1.jpg


This gene was cloned into the vector pSB1C3. PCR method was used to identify. Fig.1 shows the result of identification.

K3867001-2.jpg

Fig.1. The result of ADG1 gene cloning. M: Marker; 1: plasmid of pSB1C3-ADG1; 2: PCR result of ADG1 gene.

When this gene was constructed to the expression vector pETDuet-1, ADG1 protein was expressed. His tag was used to purify ADG1 protein. The identification result was showed in Fig.2

K3867001-3.jpg



Fig.2. The expression of ADG1 with vector pETDuet-1-ADG1 in E. coli. M, Marker; 1, Purification of ADG1 after Induced by IPTG; 2, Induced by IPTG; 3, No induced by IPTG.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 256
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 413
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]