mASIC1a cDNA, cloned between the BamHI and SacI restriction sites of the pSP64 vector, was a gift from Dr. Marcelo Carattino (University of Pittsburgh, Pittsburgh, PA). hENaC α, β, and γ subunits in the pTNT vector were a gift from Dr. Diego Alvarez de la Rosa (University of La Laguna, San Cristóbal de La Laguna, Spain). The α subunit contained the T334 and A663 polymorphisms. The construct referred to as WT hENaC contains C-terminal truncations in the β and γ subunits (β_R566STOP and γ_K576STOP) to increase expression. Site-directed mutagenesis was performed using custom-designed primers (Eurofins Genomics) and regular PCR with PfuUltra II Fusion HS DNA Polymerase (Agilent Technologies). Plasmids were linearized with EcoRI (for mASIC1a) and BamHI (for hENaC) and used as a template for synthesis of mRNA with the Ambion mMESSAGE mMACHINE SP6 Transcription Kit (Thermo Fisher Scientific). ENaCα-mASIC1aM1 chimera DNA cloned between the NheI and Xhol restriction sites in the pUNIV vector was purchased from Twist Bioscience.
Enacα masic1am1 chimera dna
Manufactured by Twist Bioscience
The ENaCα-mASIC1aM1 chimera DNA is a laboratory research tool that combines the genetic characteristics of the ENaCα and mASIC1aM1 proteins. This chimeric DNA construct allows for the expression and study of the resulting hybrid protein in experimental settings. The core function of this product is to enable researchers to investigate the structural and functional properties of this engineered protein.
Automatically generated - may contain errors
2 protocols using enacα masic1am1 chimera dna
1
Heterologous Expression of ENaC and ASIC1a
2
Chimeric ENaC-ASIC1a Channel Expression
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Mouse ASIC1a (mASIC1a) cDNA, cloned between BamHI and SacI restriction sites of the pSP64 vector, was a gift from Dr. Marcelo Carattino (University of Pittsburgh, US). hENaC a, β and γ subunits in the pTNT vector were a gift from Dr. Diego Alvarez de la Rosa (University of La Laguna, Spain). The a subunit contained the T334 and A663 polymorphisms. The construct referred to as WT hENaC contains C-terminal truncations in the β and γ subunits (β_R566STOP and γ_K576STOP) to increase expression. Site-directed mutagenesis was performed using customdesigned primers (Eurofins Genomics) and regular PCR with PfuUltra II Fusion HS DNA Polymerase (Agilent Technologies). Plasmids were linearized using EcoRI (for mASIC1a) and BamHI (for hENaC) and used as template for synthesis of mRNA with the Ambion mMESSAGE mMACHINE SP6 transcription kit (Thermo Scientific). ENaCα-mASIC1aM1 chimera DNA cloned between the NheI and Xhol restriction sites in the pUNIV vector was purchased at Twist Bioscience.
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