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Team:Peking/Texttest
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HOME
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NOTEBOOK
HUMAN PRACTICE
BIOSAFETY
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Biosensor
Bioabsorbent
Application
Expansion
Personal Notes
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Characterization
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Peking University
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Overview of the Heavy Metal Decontamination Kit
Biosensor
Introduction
Promoter Characterization
Operation Characterization
Bioreporter
Bioabsorbent
Introduction
MBP construction
MBP Expression
Facilitation Module
Inductive Aggregation
Module Assembly
Application
Preservation Optimization
Kit Operation
Expansion
Introduction
Lead Bioabsorbent
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nbsp; Secondly, we conducted a closer look into the structure of MerR family TFs(Brown et al., 2003; Hobman, 2007; Hobman et al., 2005). The metal binding domain of MerR is a 48-residue, named alpha-helix 5(Guo et al.; Song et al., 2007). A strategy to engineer MerR TF proteins was developed from previous work (Qin et al., 2006; Song et al., 2004). We directly fused two copies of alpha helix 5 into a single-chain,antiparallel coiled coil, called Metal Binding Domain (MBD). MBD reduces cost of metal binding. And then high-performance bioabsorbent was accomplished by inductively expressing MBD on surface, periplasm and cytosol of E.coli cells. The following function test results showed that our bacteria can absorb more than 95% of 10-7 mol/L Hg (II) in 5 minutes; in consistent with previous work that artificial MBD chain can simulate the in vivo metal-binding ability of dimeric, full-length MerR.
Additionally, we noticed that MerR family TFs share a high homology at their metal binding domains (Brown et al., 2003; Hobman, 2007), which implies that our strategies of bioreporter and bioabsorbent engineering might be applicable to other cases. We then expanded our reverse engineering strategy to another common toxic heavy metal, lead (Pb)(Borremans et al., 2001; Chakraborty et al., 2008; Chen et al., 2005; Julian et al., 2009; Mergeay et al., 2003). We primarily took system (lead resistance operon) apart, followed by description of function, structure and operation of PbrR, a lead responsive regulator. Then modeling was conducted to analyze the characteristics of PbrR and topology of its regulation behavior. Knowledge from the analyses confirmed us the possibility to design
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