Selected article for: "cdna fragment and GenBank accession"
Author: Yu, Liping; Zhang, Xiaorong; Wu, Tianqi; Su, Jin; Wang, Yuyang; Wang, Yuexin; Ruan, Baoyang; Niu, Xiaosai; Wu, Yantao
Title: Avian infectious bronchitis virus disrupts the melanoma differentiation associated gene 5 (MDA5) signaling pathway by cleavage of the adaptor protein MAVS
  • Document date: 2017_11_13
    • ID: 0zn1sqj9_7
    Snippet: The chMDA5 ORF was amplified from CEK cells by overlap PCR with primers chMDA5-F1/R1 and chMDA5-F2/R2, producing a 3006 bp MDA5 PCR product (GenBank accession No. GU570144.1). The PCR product was digested with EcoR V and Xba I, then was inserted into the pcDNA-5′-Flag plasmid that had been digested with the same enzymes. The chTLR3 ORF was PCR amplified from CEK cells with the primer pair chTLR3 F/R, and the product was cloned into the p3 × fl.....
    Document: The chMDA5 ORF was amplified from CEK cells by overlap PCR with primers chMDA5-F1/R1 and chMDA5-F2/R2, producing a 3006 bp MDA5 PCR product (GenBank accession No. GU570144.1). The PCR product was digested with EcoR V and Xba I, then was inserted into the pcDNA-5′-Flag plasmid that had been digested with the same enzymes. The chTLR3 ORF was PCR amplified from CEK cells with the primer pair chTLR3 F/R, and the product was cloned into the p3 × flag-CMV-7.1 vector (Invitrogen) at the Not I and EcoR V sites. The dgRIG-I ORF (GenBank accession No. JF804977) was amplified from goose splenic cDNA using primers dgRIG-I F/R, and the fragment was inserted into the p3 × flag-CMV-7.1 vector at the EcoR I and BamH I sites. The small interfering RNAs (siRNA) targeting the chicken chMDA5, chTLR3, and chMAVS mRNAs as well as control siRNA were synthesized by Santa Cruz Biotechnology and have been previously described [22] .

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