Description
P
alcA is one of the strongest inducible promoters in
Aspergillus nidulans commonly used to overexpress proteins [1]. It has been shown that P
alcA promoter is also functional in monocotyledonous plant sugar cane [2] and
Escherichia coli (E.coli)[3]. Its transcriptional activation is dependent on the binding of its positive transcriptional regulator AlcR with various substrates that employ a hydroxyl group, for example ethanol and threonine. The native P
alcA consists of 3 AlcR binding sites. The number and position of the AlcR binding sites on the P
alcA are crucial in determining its transcriptional activation strength. It has also been shown that each AlcR target in the P
alcA contributes differently to the activation of the downstream protein expression [1].
This part is a variant of P
alcA (BBa_K2092002). In contrast to the native construct (three AlcR binding sites
abc), this variant consists only two binding sites
bc. Previous study on sugarcane has shown that there is a 70% decrease in promoter strength for P
alcA with only binding sites
bc [2]. This finding, however, has not been verified using
E.coli as a chassis. This then forms the very basis of our project.
References:
[1] Panozzo, C., Capuano, V., Fillinger, S. and Felenbok, B. (1997). The zinc binuclear cluster Activator AlcR is able to bind to single sites but requires multiple repeated sites for synergistic activation of the alcA gene in
Aspergillus nidulans.
Journal of Biological Chemistry, 272(36), 22859???22865.
[2] Kinkema, M., Geijskes, R.J., Shand, K., Coleman, H.D., De Lucca, P.C., Palupe, A., Harrison, M.D., Jepson, I., Dale, J.L. and Sainz, M.B. (2013). An improved chemically inducible gene switch that functions in the monocotyledonous plant sugar cane.
Plant Molecular Biology, 84(4-5), 443???454.
[3] Hemmati, H. and Basu, C. (2015). Transcriptional analyses of an ethanol inducible promoter in
Escherichia coli and tobacco for production of cellulase and green fluorescent protein.
Biotechnology & Biotechnological Equipment, 29(6), 1043???1052.