Part:BBa_K4288010
ArsA-GFP
ArsA-GFP
Profile
Name: pro-ArsA amilGFP
Base Pairs: 2564 bp
Origin: Escherichia coli
Properties: Gene technology for protecting patented bacterial strains
Usage and Biology
ArsA was designed to response to the various concentration of arsenic, and fused amilGFP to monitor the arsenic concentration.
BBa_K3991000
Name: ArsA
Base Pairs: 1749 bp
Origin: Escherichia coli
Properties: arsenic metallochaperone
Usage and Biology
The ArsA protein is an arsenite-stimulated ATPase and complexed with ArsB protein. Its function is to transport the arsenic.
Bacteria developed a mechanism against the arsenic pervasiveness. Many bacteria processed three genes, arsRBC. Five gene ars operons have two additional genes, arsD and arsA, called arsRDABC. ArsR is an As(III)-responsive transcriptional repressor, additional genes ArsD and arsA derived from E.coli. The arsRDABC operon are more resistant to As due to the ArsA-ArsB complex that catalyzes ATP-driven As/Sb efflux.
Construct design
In order to develop a real-time tool for detecting the arsenic binding, promotor ArsA was designed to response to the various concentration of arsenic, fused to amilGFP to monitor the arsenic concentration. This DNA fragment was inserted into the expression vector pET28a.
Experimental approach
The result demonstrated the relationship between the florescence intensity and the arsenic concentration ranging from 10ug/L to 200ug/L. Compared to cultivation time of 1h or 0h, the green curve of cultivation time of 2h showed the significant increasing GFP intensity. however, the higher concentration of arsenic (100ug/L) might inhibit the bacteria growth, so the GFP intensity decreased. According the result, 50ug/L induced the maximum florescence expression under ArsA promoter.
Reference
1. Silver, S. and L.T. Phung, BACTERIAL HEAVY METAL RESISTANCE: New Surprises.[J] Annual Review of Microbiology, 1996. 50(1):753-789.
2. Lin, Y.-F., J. Yang, and B.P. Rosen, ArsD: an As(III) metallochaperone for the ArsAB As(III)-translocating ATPase.[J] Journal of Bioenergetics and Biomembranes, 2007. 39(5):453-458.
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