Method for improving yield of homoorientin produced by recombinant bacteria through coordinated regulation strategy
A technology of isoorientin and recombinant bacteria, applied in the fields of genetic engineering technology and microorganisms, can solve the problems such as the inability to greatly increase the yield, and achieve the effects of inhibiting the accumulation of acetic acid, increasing the yield and simple process
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[0021] 1. Construction of recombinant bacteria BL21-CGT-I
[0022] The carboside glycosyltransferase Gt6CGT gene (AB985754.1) derived from Gentiana triflora was optimized according to the dominant codon table of Escherichia coli K12 (http: / / www.kazusa.or.jp / codon / ), and the amino acid The codons with high frequency of sense codons are used as dominant codons, and finally the optimized sequence information of the carboside glycosyltransferase gene is obtained, which is synthesized by Shanghai Jierui Bioengineering Co., Ltd. and added to the N-terminus and C-terminus of the gene respectively The Nco I and BamH I restriction sites were used to obtain the optimized Gt6CGT gene, the gene sequence of which is shown in SEQ ID NO:1.
[0023] The optimized Gt6CGT gene was double-digested with Nco I and BamH I, and the recombinant plasmid pCDFDuet-1 was also double-digested with Nco I and BamH I, and the rubber was harvested separately, concentrated and ligated overnight at 16°C, and th...
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