Selected article for: "chain reaction and reading frame"
Author: Tang, Hei-Man Vincent; Gao, Wei-Wei; Chan, Chi-Ping; Cheng, Yun; Chaudhary, Vidyanath; Deng, Jian-Jun; Yuen, Kit-San; Wong, Chun-Ming; Ng, Irene Oi-Lin; Kok, Kin-Hang; Zhou, Jie; Jin, Dong-Yan
Title: Requirement of CRTC1 coactivator for hepatitis B virus transcription
  • Document date: 2014_11_10
    • ID: qtoygz6w_12
    Snippet: Reporter plasmids preS2-Luc, X-Luc, preS1-Luc and C-Luc were derived from pGL3-Basic (Promega). preS1 (nucleotides 2710-2834), preS2 (nucleotides 2966-152), X (nucleotides 950-1310) and C (nucleotides 1403-1817) promoter sequences were amplified by polymerase chain reaction (PCR) from the HBV genome of adw2 subtype (Genbank AM282986) in plasmid pHBV1.3D, which has been described elsewhere (9, 23) . Expression plasmids for CREB, A-CREB, CRTC1, CRT.....
    Document: Reporter plasmids preS2-Luc, X-Luc, preS1-Luc and C-Luc were derived from pGL3-Basic (Promega). preS1 (nucleotides 2710-2834), preS2 (nucleotides 2966-152), X (nucleotides 950-1310) and C (nucleotides 1403-1817) promoter sequences were amplified by polymerase chain reaction (PCR) from the HBV genome of adw2 subtype (Genbank AM282986) in plasmid pHBV1.3D, which has been described elsewhere (9, 23) . Expression plasmids for CREB, A-CREB, CRTC1, CRTC2, CRTC3, CRTC1-S167A and CRTC1-M1, as well as reporter plasmid pCRE-Luc have also been detailed previously (15, 24) . The expression plasmid for HBx was based on the pCAGEN vector and the HBx open reading frame (nucleotides 1374-1838) was PCR amplified from pHBV1.3D with the addition of a Flag-HA tag to its 5 end. The X-deficient pHBV1.3D was made by converting the eighth codon of HBx to a stop codon. The mutagenic primers were 5 -CGCGAAGGAT CCAGT-TAGCA GTACAACCTA GCA-3 and 5 -GCGCTTCCTA GGTCAATCGT CATGTTGGAT CGT-3 . A similar construct has been described elsewhere (25) . Expression vector for the lysine-free (K0) mutant of ubiquitin is a kind gift from Dr James Chen (26) .

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