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Vector Information
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Size (kb): 5.3000001907348630 Vector: pETGEXCT (plasmid) Promoters: Promoter T7 (phi10) Construction: pET3d, pGEX2T Marker(s):ampR Construct size (kb): 5.300000190734863 Features: marker(s): ampR
other: thrombin cleavage site I
other: thrombin cleavage site II
promoter: T7 (phi10)
replicon: pMB1
restriction site: BamHI
restriction site: NcoI
restriction site: SacI
coding sequence: GST
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Applications
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encodes removable tag for protein isolation expression vector vector permitting RNA synthesis in vitro vector permitting construction of fusion proteins
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Comments
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Restriction digests of the clone give the following sizes (kb): BamHI--5.6; NcoI--5.6; SstI--5.6. The thrombin cleavage sites produced in the protein enable the release of the fused protein from the GST tag. The order of the major features in the plasmid is: T7 promoter - NcoI - SacI - thrombin I encoding site - GST coding sequence - thrombin II encoding site - BamHI - ampR - pMB1 ori. The plasmid yield is rather low and thrombin cleavage of the fusion proteins can be inefficient. Expression vector allowing production of an N-terminal or C-terminal fusion to glutathione S-transferase (GST) for affinity purification of the recombinant protein.
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References
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Sharrocks AD. A T7 expression vector for producing N- and C-terminal fusion proteins with glutathione S-transferase. Gene 138: 105-108, 1994. PubMed: 8125285
Andrew D Sharrocks, personal communication
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