Difference between revisions of "Part:BBa K3867007"
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Fig.1. The result of GBSS1 gene cloning. | Fig.1. The result of GBSS1 gene cloning. | ||
M: Marker; 1: plasmid of pSB1C3-GBSS1; 2: PCR result of GBSS1 gene. | M: Marker; 1: plasmid of pSB1C3-GBSS1; 2: PCR result of GBSS1 gene. |
Latest revision as of 09:19, 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.
This 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 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
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1465