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<h2>2.Express and verify RdhANP</h2> | <h2>2.Express and verify RdhANP</h2> | ||
<h3>2.1 Clone rdhANP to suitable vectors </h3> | <h3>2.1 Clone rdhANP to suitable vectors </h3> | ||
− | + | <h3>2.2 Induce to express RdhANP in B. megaterium</h3> | |
− | + | <h3>2.3 Purify and concentrate RdhANP </h3> | |
− | + | <h3>2.4 Construct dehalogenation-testing system in vitro and in vivo</h3> | |
− | + | <h3>2.5 Detect dehalogenation reaction by HPLC-MS</h3> | |
− | + | <h3>2.6 Overexpress RdhANP by testing different vectors </h3> | |
− | + | <h3>2.7 Design synthetic RBS by 'Ribosome binding site calculator' and RBSDesigner and test their production level</h3> | |
− | + | <h3>2.8 Perform Directed Enzyme& Protein Evolution by certain companies. </h3> | |
− | + | <h3>2.9 Test the dehalogenation of these mutants our constructed wastewater-imitating system </h3> | |
− | + | <h3>2.10 Coordinate B12 production and RdhANP expression</h3> | |
<h2>3.Bind B. megaterium to cellulose membrane</h2> | <h2>3.Bind B. megaterium to cellulose membrane</h2> | ||
− | 3.1 Coexpress Cellulose binding domain(CBD) and Membrane anchoring domain(MAD) in B. megaterium | + | <h3>3.1 Coexpress Cellulose binding domain(CBD) and Membrane anchoring domain(MAD) in B. megaterium</h3> |
− | 3.2 Test the binding of B. megaterium to cellulose membrane | + | <h3>3.2 Test the binding of B. megaterium to cellulose membrane</h3> |
<h2>4.Coexpress CBD, MAD and RdhANP in B. megaterium and test its binding to cellulose membrane and dehalogenation</h2> | <h2>4.Coexpress CBD, MAD and RdhANP in B. megaterium and test its binding to cellulose membrane and dehalogenation</h2> | ||
<h2>5.Construct biofilm containing RdhANP-expressing B. megaterium</h2> | <h2>5.Construct biofilm containing RdhANP-expressing B. megaterium</h2> |
Revision as of 07:09, 23 October 2017
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