抄録
Engineering and design of genetic circuit in living cell is critical in accessing the beneficial application of synthetic biology. Directed evolution can avoid the complicated rational design of such circuit by screening or selecting functional circuit from non-functional one. Here, we proposed a positive-negative selection system for selecting a transcription factor that activates gene expression in response to arsenic in solution. First, we developed a whole cell biosensor for sensing arsenite in liquid using a regulator (ArsR) and a reporter (GFP), and evaluated its performance. Second, we developed a positive selection system for active ArsR using compartmentalized partnered replication that uses thermostable DNA polymerase as the reporter of activity. Third, we developed a negative selection system using sucrose-induced suicide gene SacB as the reporter for exclusion of inactive ArsR variants.
| 元の言語 | English |
|---|---|
| ページ（範囲） | 341-348 |
| ページ数 | 8 |
| ジャーナル | Journal of Biochemistry |
| 巻 | 164 |
| 発行部数 | 5 |
| DOI | |
| 出版物ステータス | Published - 2018 11 1 |
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ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
これを引用
A dual system using compartmentalized partnered replication for selection of arsenic-responsive transcriptional regulator. / Aye, Seaim Lwin; Fujiwara, Kei; Doi, Nobuhide.
：: Journal of Biochemistry, 巻 164, 番号 5, 01.11.2018, p. 341-348.研究成果: Article
}
TY - JOUR
T1 - A dual system using compartmentalized partnered replication for selection of arsenic-responsive transcriptional regulator
AU - Aye, Seaim Lwin
AU - Fujiwara, Kei
AU - Doi, Nobuhide
PY - 2018/11/1
Y1 - 2018/11/1
N2 - Engineering and design of genetic circuit in living cell is critical in accessing the beneficial application of synthetic biology. Directed evolution can avoid the complicated rational design of such circuit by screening or selecting functional circuit from non-functional one. Here, we proposed a positive-negative selection system for selecting a transcription factor that activates gene expression in response to arsenic in solution. First, we developed a whole cell biosensor for sensing arsenite in liquid using a regulator (ArsR) and a reporter (GFP), and evaluated its performance. Second, we developed a positive selection system for active ArsR using compartmentalized partnered replication that uses thermostable DNA polymerase as the reporter of activity. Third, we developed a negative selection system using sucrose-induced suicide gene SacB as the reporter for exclusion of inactive ArsR variants.
AB - Engineering and design of genetic circuit in living cell is critical in accessing the beneficial application of synthetic biology. Directed evolution can avoid the complicated rational design of such circuit by screening or selecting functional circuit from non-functional one. Here, we proposed a positive-negative selection system for selecting a transcription factor that activates gene expression in response to arsenic in solution. First, we developed a whole cell biosensor for sensing arsenite in liquid using a regulator (ArsR) and a reporter (GFP), and evaluated its performance. Second, we developed a positive selection system for active ArsR using compartmentalized partnered replication that uses thermostable DNA polymerase as the reporter of activity. Third, we developed a negative selection system using sucrose-induced suicide gene SacB as the reporter for exclusion of inactive ArsR variants.
UR - http://www.scopus.com/inward/record.url?scp=85054892027&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85054892027&partnerID=8YFLogxK
U2 - 10.1093/jb/mvy055
DO - 10.1093/jb/mvy055
M3 - Article
C2 - 29893865
AN - SCOPUS:85054892027
VL - 164
SP - 341
EP - 348
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 5
ER -