Materials and Reagents

1. Bacillus cereus HYC-7
   
2. Escherichia coli (E.coli) DH5α strain, bacteriophage T7
   
3. Glyphosate (Sigma-Aldrich, catalog number: PS1051 )
   
4. Tryptone (Difco)
   
5. Yeast extract (Difco)
   
6. Ampicillin
   
7. o-Dianisidine dihydrochloride (Sigma-Aldrich, catalog number: D3252 )
   
8. Horseradish peroxidase (Sigma-Aldrich, catalog number: P6782 )
   
9. Protein expression vector of pGEX-6P-1 (the plasmid full length 4,984 bp) (GE Healthcare, catalog number: 28-9546-48 ; Genbank accession number: U78872.1)
   
10. Recombinant plasmid pGEX-GO contains encoding gene of glycine oxidase from Bacillus cereus HYC-7
    The nucleotide sequence (1,110 bp) was submitted to the NCBI Genbank and gained the accession number (KC203486.1).
    
11. Taq DNA polymerase (Takara, catalog number: R500A )
    
12. DpnI restriction enzyme (Takara, catalog number: 1235A )
    
13. dATP, dTTP, dCTP, dGTP (Takara, catalog numbers: 4026Q , 4029Q , 4028Q , 4027Q )
    
14. TransStart^® FastPfu DNA polymerase (TransGen Biotech, catalog number: AP221-01 )
    
15. High Pure dNTPs (TransGen Biotech, catalog number: AD101-01 )
    
16. Luria-Bertani medium (see Recipes)
    

Equipment

1. 96 deep-well plates (Axygen, catalog number: P-DW-20-C-S )
   
2. Gel purification column (Axygen)
   
3. Thermo Multiskan spectrum plate reader (Thermo Scientific, catalog number: 51118600 )
   
4. Thermal cyclers (Bio-Rad Laboratories, catalog number: 186-1096 )
   
5. Ultrasonic processor (Sigma-Aldrich, catalog number: Z412619-1EA )
   

Procedure