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pSGP18 BioVector?畢赤酵母CBP和His標簽表達載體 BioVector NTCC質粒載體菌株細胞蛋白抗體基因保藏中心

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pSGP18 BioVector?畢赤酵母CBP和His標簽表達載體

BioVector NTCC質粒載體菌株細胞蛋白抗體基因保藏中心


pSGP18 is a yeast expression vector. Here's a breakdown of its key features:

  • Yeast Expression: It's designed for protein expression in the yeast Pichia pastoris.

  • AOX1 Promoter: This promoter drives strong gene expression in Pichia pastoris when induced with methanol.

  • C-terminal Tags:

    • CBP (Calmodulin Binding Protein): Allows for affinity purification of the expressed protein using calmodulin affinity chromatography.

    • 6xHis-tag: Enables protein purification using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography.

    • 3C Protease Cleavage Site: Allows for the removal of the CBP and 6xHis-tags after purification.


  • Zeocin Resistance: Confers resistance to the antibiotic zeocin, allowing for selection of yeast cells that have successfully integrated the plasmid.

  • Ligation-Independent Cloning (LIC): This method simplifies the process of inserting genes into the vector.

Key Applications:

  • Heterologous Protein Expression: pSGP18 is widely used to express recombinant proteins in Pichia pastoris. This yeast system offers several advantages, including high protein yields, proper protein folding and post-translational modifications, and ease of cultivation.

  • Protein Purification: The CBP and 6xHis-tags facilitate efficient purification of the expressed protein using affinity chromatography.

  • Protein Characterization: The expressed protein can be further characterized for its biochemical and functional properties.

Vector Name:
pSGP18
Synonyms:
''
Sequencing Primer:
Forward:pSGP18for
Reverse:  Unknown
Description:
Yeast expression vector with a AOX1 promoter C-terminal CBP and RSG-6xHis tag with 3C protease cleavage site; zeocin resistance in Pichia and in bacteria; ligation independent cloning (LIC)
Comments:
Similar to pSGP17 but without the prepro-alpha-factor signal sequence
Size (bp):
3424
Parent Vector:
None
Properties:
expression in Pichia pastoris, ligation independent cloning (LIC), with tag/fusion/marker, yeast expression
Author Name:
University of Rochester
Mark Dumont
Sara M Connelly
Membrane Protein Structural Biology Consortium
PSI:Biology
Publications:
Title: Purification and ATP hydrolysis of the putative cholesterol transporters ABCG5 and ABCG8
Title: Expression of 25 human ABC transporters in the yeast Pichia pastoris and characterization of the purified ABCC3 ATPase activity

Vector Features:

Type
Name
Description
Start
End
bacterial origin
pUC ori
pUC origin for bacterial replication
2738
3420
ligation independent cloning
LIC
LIC cloning site (using an inverted pair of BsmBI sites with a PstI site in between)
956
975
primer
primer
forward sequencing primer CACCTGTGCCGAAACGCAAATG
625
646
promoter
AOX1
Pichia AOX1 promoter
7
935
protease cleavage site
3C
3C protease cleavage site
988
1011
selectable marker
ZeoR
Zeocin resistance gene
1994
2368
tag
His
C-terminal RGS-6xHis tag
1151
1168
tag
CBP
C-terminal calmodulin binding peptide (CBP) tag (start and end location estimated)
1012
1150

The DREAM method allows site-directed mutagenesis of an unstable... |  Download Scientific Diagram

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