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Pcdh expression vector

Manufactured by System Biosciences
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The PCDH expression vector is a plasmid designed for the expression of protocadherin (PCDH) genes in mammalian cell lines. The vector contains the necessary elements for efficient gene expression, including a promoter, multiple cloning site, and a selection marker. This product can be used to study the function and regulation of PCDH genes in various cellular and experimental contexts.

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Lab products found in correlation

Gibson assembly, by New England Biolabs (2 mentions) Kapa hifi pcr kit, by Roche (2 mentions) Pgex 4t 3, by Cytiva (2 mentions) Pgex f primer, by Cytiva (2 mentions)

4 protocols using pcdh expression vector

1

Construction of Cre-Intein Fusion Plasmids

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NCre and CCre were generated from the pCAG-Cre:GFP, a gift from Connie Cepko (Addgene plasmid #13776; https://www.addgene.org/13776/; RRID:Addgene_13,776) using the KAPA HiFi PCR kit (Roche Sequencing). IntN and IntC were amplified from the Npu DnaE inteins with site directed mutagenesis according to Lockless and Muir [18 (link)]. All plasmids were subcloned into a pCDH expression vector (System Biosciences), assembled using Gibson Assembly (New England Biolabs) and verified by Sanger DNA sequencing.
Khoo A.T., Kim P.J., Kim H.M, & Je H.S. (2020). Neural circuit analysis using a novel intersectional split intein-mediated split-Cre recombinase system. Molecular Brain, 13, 101.
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2

Cloning Alanine-tagged Constructs

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An oligonucleotide encoding a 20 Alanine peptide was cloned into pGEX4T-3 (Amersham Biosciences) to create the construct containing glutathione-S-transferase containing 20 carboxy-terminal alanine residues (GST-A20). Constructs were confirmed by dideoxynucleotide sequencing using the pGEX-F primer (Amersham Biosciences). The PABPN1 A10 (WT) and PABPN1 A13–17 constructs were cloned into the pCDH expression vector (System Biosciences, Mountain View, CA) containing a 6x-His tag (Table 1). Constructs encoding RUNX2 WT and RUNX2 A27 were cloned into the expression vector pTL1-HA, which contains an HA tag (15 (link)).
Vest K.E., Apponi L.H., Banerjee A., Pavlath G.K, & Corbett A.H. (2015). An Antibody to Detect Alanine-Expanded PABPN1: A New Tool to Study Oculopharyngeal Muscular Dystrophy. Journal of Neuromuscular Diseases, 2(4), 439-446.
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3

Cloning and Expressing Alanine-Rich Proteins

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An oligonucleotide encoding a 20 Alanine peptide was cloned into pGEX4T-3 (Amersham Biosciences) to create the construct containing glutathione-S-transferase containing 20 carboxy-terminal alanine residues (GST-A20). Constructs were confirmed by dideoxynucleotide sequencing using the pGEX-F primer (Amersham Biosciences). The PABPN1 A10 (WT) and PABPN1 A13-17 constructs were cloned into the pCDH expression vector (System Biosciences, Mountain View, CA) containing a 6x-His tag (Table 1). Constructs encoding RUNX2 WT and RUNX2 A27 were cloned into the expression vector pTL1-HA, which contains an HA tag [15 (link)].
Vest K.E., Apponi L.H., Banerjee A., Pavlath G.K, & Corbett A.H. (2015). An Antibody to Detect Alanine-Expanded PABPN1: A New Tool to Study Oculopharyngeal Muscular Dystrophy. Journal of Neuromuscular Diseases, 2(4), 439-446.
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4

Generating Versatile Cre/intein Constructs

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NCre and CCre were generated from the pCAG-Cre:GFP, a gift from Connie Cepko (Addgene plasmid #13776; https://www.addgene.org/13776/ ; RRID:Addgene_13776) using the KAPA HiFi PCR kit (Roche Sequencing). IntN and IntC were amplified from the Npu DnaE inteins with site directed mutagenesis according to Lockless and Muir (2009) . All plasmids were subcloned into a pCDH expression vector (System Biosciences), assembled using Gibson Assembly (New England Biolabs) and verified by Sanger DNA sequencing.
Khoo A., Kim P.J., Kim H.M., & Je H.S. (2020). Neural circuit analysis using a novel intersectional split intein-mediated split-Cre recombinase system.
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