TAILIEUCHUNG - Báo cáo khoa học: Photoregulation of DNA transcription by using photoresponsive T7 promoters and clarification of its mechanism

With the use of photoresponsive T7 promoters tethering two 2¢-methylazo-benzenes or 2¢,6¢-dimethylazobenzenes, highly efficient photoregulation of DNA transcription was obtained. After UV-A light irradiation (320–400 nm), the rate of transcription with T7 RNA polymerase and a photoresponsive promoter involving two 2¢,6¢-dimethylazobenzenes was 10-fold faster than that after visible light irradiation (400–600 nm). | ỊFEBS Journal Photoregulation of DNA transcription by using photoresponsive T7 promoters and clarification of its mechanism Xingguo Liang1 Ryuji Wakuda1 Kenta Fujioka1 and Hiroyuki Asanuma1 2 1 Department of Molecular Design and Engineering Graduate Schoolof Engineering Nagoya University Japan 2 CREST Japan Science and Technology Agency JST Kawaguchi Japan Keywords azobenzene modified DNA photoregulation T7 promoter transcription Correspondence X. G. Liang Department of Molecular Design and Engineering Graduate School of Engineering Nagoya University Chikusa-ku Nagoya 464-8603 Japan Fax 81 52 789 2528 Tel 81 52 789 2488 E-mail liang@ H. Asanuma Department of Molecular Design and Engineering Graduate School of Engineering Nagoya University Chikusa-ku Nagoya 464-8603 Japan Fax 81 52 789 2528 Tel 81 52 789 2488 E-mail asanuma@ Received 10 December 2009 revised 11 January 2010 accepted 15 January 2010 With the use of photoresponsive T7 promoters tethering two 2 -methylazo-benzenes or 2 6 -dimethylazobenzenes highly efficient photoregulation of DNA transcription was obtained. After UV-A light irradiation 320-400 nm the rate of transcription with T7 RNA polymerase and a photoresponsive promoter involving two 2 6 -dimethylazobenzenes was 10-fold faster than that after visible light irradiation 400-600 nm . By attaching a nonmodified azobenzene and 2 6 -dimethylazobenzene at the two positions respectively and by utilizing the different cis fi trans thermal stability between cis-nonmodified azobenzene and cis-2 6 -dimethy-lazobenzene four species of T7 promoter cis-cis trans-cis cis-trans and trans-trans were obtained. The four species showed transcriptional activity in the order of cis-cis cis-trans trans-cis trans-trans. Kinetic analysis revealed that the Km for the cis-cis promoter both of the introduced azobenzene derivatives were in the cis form and T7 RNA polymerase was 68 times lower than that for the trans-trans form indicating .

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