Part:BBa_K1023001

pkiA promotor

xlnD terminator direction Forward forward_efficiency -NA- reversed_version -NA- reverse_efficiency -NA-

The Aspergillus niger pyruvate kinase gene was isolated and characterized by De Graaff et al. 1992. The pkiA promoter of A. niger is constitutive and has a high expression level. Target genes can be expressed under the control of this highly efficient promoter (Roth, A. H., & Dersch, P. 2010). In our project, we first introduced the pkiA promoter into iGEM to express the codon optimized eforRed encoding gene with signal sequence, with which we succesfully transformed A. niger N593. The mycelium of the A. niger transformants displayed the red colour and it proved the pkiA could express the chromoprotein correctly. (see Fig. 1). Besides, the successful expression of ATP sensor also confirmed that the pkiA promoter could express inserted genes efficiently in A. niger. (see Fig. 2)

Fig.1 Transformant (left) A. niger proved that the pkiA promotor was functional as red mycelium was clearly visible. The negative control (non codon-optimized eforRed) didn't have any colour.

Fig.2 ATP biosensor expression in A.niger

Sequence and Features BBa_K1023001 ===References=== 1. de Graaff, L., van den Broeck, H., & Visser, J. (1992). Isolation and characterization of the Aspergillus niger pyruvate kinase gene. Current genetics, 22(1), 21-27. 2. Roth, A. H., & Dersch, P. (2010). A novel expression system for intracellular production and purification of recombinant affinity-tagged proteins in Aspergillus niger. Applied microbiology and biotechnology, 86(2), 659-670.