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Improvement Parts

Name	Number	Type	Description	Length
lldRO1-PfnrF8-lldRO2	BBa_K2913023	Promoter	Response to high lactate and hypoxia. This is a composite part consists of lldRO1-J23117-lldRO2 (BBa_K1847007) and PfnrF8 (BBa_K2913009). It has a functional improvement of lldRO1-J23117-lldRO2 via transforming a specific Anderson promoter (BBa_J23117) to a hypoxia-inducible promoter, PfnrF8. The lldRO1--lldRO2 was utilized with other parts of the lldPRD operon (previously named as lct), responsible for aerobic L-lactate metabolism. The lldPRD operon consists of three genes that form a single transcriptional unit inducible by growth in L-lactate. The three genes lldD, lldP and lldR encode a dehydrogenase, a permease and a regulatory protein, respectively. We chose lldR and lldP to assist building our lactic acid response unit. lldR protein can bind to the operators O1 and O2 located on each side of the PfnrF8. When lactic acid level is low, two lldR molecules will individually bind to the O1 and O2 sites and form a tetramer to make DNA strand form a hairpin structure, which can turn off the expression of the downstream gene. When lactic acid level reaches a certain point, lldR will be released from the O1 and O2 operators, and the DNA hairpin will be resolved, leading to activated transcription of the downstream gene. We also used lldP to increase the sensitivity of our system to lactic acid alteration.	159 bp

Team:NEFU China/Improve

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