A kind of amino acid racemase and its application
An amino acid, leucine technology, applied in the direction of racemase/epimerase, enzyme, isomerase, etc., can solve problems such as narrow substrate spectrum
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[0030] 1. Construction of recombinant expression vector
[0031] The coding gene TkAAR (nucleotide sequence shown in SEQ ID NO.2) was connected to the cloning vector pUC118, and the connection product was transformed into E. coli Escherichia coli DH5α competent cells, and spread on the LB plate culture medium containing ampicillin . After picking a single colony, insert it into LB liquid medium for culture and extract the plasmid for sequencing. The one with the correct sequencing result is the cloning vector pUC118-TK1211.
[0032] The recombinant cloning vector pUC118-TK1211 and the ESS00988 vector containing a strong promoter Pcsg were treated with NdeI and SalI double digestion, and the target gene and the vector after double digestion were ligated overnight at 16°C. The ligation product was transformed into Escherichia coli E.coli DH5α competent cells, and spread on LB plate medium containing ampicillin after recovery. Pick a single colony, insert it into LB liquid medi...
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