Difference between revisions of "Part:BBa K3867007"

 
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GBSS1 (Granule-bound starch synthase1) is another key enzyme for the amylose synthesis. It belongs to UDP-Glycosyltransferase superfamily protein. GBSS1 is an enzyme that catalyzes the transfer of glucose from to glucose-containing polysaccharides with α1,4-linkages, which is the main gene controlling amylose synthesis.
 
GBSS1 (Granule-bound starch synthase1) is another key enzyme for the amylose synthesis. It belongs to UDP-Glycosyltransferase superfamily protein. GBSS1 is an enzyme that catalyzes the transfer of glucose from to glucose-containing polysaccharides with α1,4-linkages, which is the main gene controlling amylose synthesis.
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[[File:K3867003-1.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:K3867002-2.jpg|center]]
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Fig.1. The result of GBSS1 gene cloning.
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M: Marker; 1: plasmid of pSB1C3-GBSS1; 2: PCR result of GBSS1 gene.
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When GBSS1 gene was constructed to the expression vector pET-28a(+), GBSS1 protein was expressed. His tag was used to purify GBSS1 protein. The identification result was showed in Fig.2
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[[File:K3867003-3.jpg|center]]
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Fig.2. The expression of GBSS1 with vector pET-28a(+)-GBSS1 in E. coli.
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M, Marker; 1.No induced by IPTG; 2.Induced by IPTG; 3.Purification of GBSS1 after induced by IPTG; 
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Revision as of 09:18, 14 October 2021


GBSS1 (Granule-bound starch synthase1)

GBSS1 (Granule-bound starch synthase1) is another key enzyme for the amylose synthesis. It belongs to UDP-Glycosyltransferase superfamily protein. GBSS1 is an enzyme that catalyzes the transfer of glucose from to glucose-containing polysaccharides with α1,4-linkages, which is the main gene controlling amylose synthesis.

K3867003-1.jpg


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

K3867002-2.jpg

Fig.1. The result of GBSS1 gene cloning. M: Marker; 1: plasmid of pSB1C3-GBSS1; 2: PCR result of GBSS1 gene.

When GBSS1 gene was constructed to the expression vector pET-28a(+), GBSS1 protein was expressed. His tag was used to purify GBSS1 protein. The identification result was showed in Fig.2

K3867003-3.jpg



Fig.2. The expression of GBSS1 with vector pET-28a(+)-GBSS1 in E. coli. M, Marker; 1.No induced by IPTG; 2.Induced by IPTG; 3.Purification of GBSS1 after induced by IPTG;


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 163
    Illegal BglII site found at 202
    Illegal BamHI site found at 538
    Illegal BamHI site found at 1599
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1465