Difference between revisions of "Part:BBa K3867001"

 
 
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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.  
 
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.  
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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.
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[[File:K3867001-2.jpg|center]]
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Fig.1. The result of ADG1 gene cloning.
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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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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]