Difference between revisions of "Part:BBa K3867005"

Revision as of 08:43, 14 October 2021

APL1 (ADP glucose pyrophosphorylase large subunit 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.

APL1 gene was cloned into the vector pSB1C3. PCR method was used to identify. Fig.1 shows the result of identification.

Fig.1. The result of APL1 gene cloning. M: Marker; 1: plasmid of pSB1C3-APL1; 2: PCR result of APL1 gene.

When APL1 gene was constructed to the expression vector pETDuet-1, APL1 protein was expressed. His tag was used to purify APL1 protein. The identification result was showed in Fig.2

Fig.2. The expression of APL1 with vector pETDuet-1-APL1 in E.coli. M, Marker; 1, No induced by IPTG; 2, Purification of APL1 protein (57.5kD) induced by IPTG; 3, induced by IPTG.

Sequence and Features