Part:BBa_K4968017
CBM-sfGFP-pUC_GW_KanR-SUMO
The component part consist of SmtA (BBa_K4968001), CBM (BBa_K4968002), sfGFP (BBa_K4968003)
SmtA (BBa_K4968001) is a protein belonging to the metallothionein family, capable of adsorbing heavy metals and chelating them in an inactive form. By introducing the carbohydrate-binding module (CBM) (BBa_K4968002), our goal is to tightly bind the protein to cellulose, thereby creating a bioremediation agent with potential applications. The addition of superfolder green fluorescent protein (sfGFP) (BBa_K4968003) is used to confirm protein expression, monitor the immobilization process, and study protein dynamics.
Usage & Biology
Cyanobacterial metallothionein SmtA (BBa_K4968001), originating from Synechococcus PCC 7942, is a protein belonging to the metallothionein family. It can sense and bind specific heavy metal ions such as cadmium, lead, zinc, etc., thereby assisting in regulating the cell's tolerance and metabolism of these toxic metals. It prevents these metal ions from entering the cell or transferring them to cellular compartments by forming complexes with them, thus protecting the cell from the toxicity of heavy metals.
Carbohydrate-binding modules (CBMs) (BBa_K4968002)are components of several enzymes that can bind to specific carbohydrates. One of their main functions is to be used in constructing specific bifunctional proteins (Oliveira et al., 2015). CBMs are capable of recognizing and selectively binding to characteristics on the crystal surface, providing possibilities for modifying cellulose. By introducing carbohydrate-binding modules (CBMs), our goal is to tightly attach proteins to cellulose, thereby creating a bioadsorbent with potential applications.
sfGFP (BBa_K4968003) is a fluorescent labeling tool that, compared to other GFP variants, does not induce misfolding of fusion proteins when expressed alongside other proteins (Pédelacq et al., 2005), enhancing the stability of the fusion protein. Additionally, sfGFP exhibits superior performance and is suitable for protein-protein interactions, monitoring immobilization processes, and expression studies.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 3885
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 3885
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 3885
Illegal XhoI site found at 1078 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 3885
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 3885
Illegal AgeI site found at 487 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 93
Illegal BsaI site found at 3760
Illegal BsaI.rc site found at 86
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