Method for degrading plant lignin by using Bacillus subtilis engineering bacterial
A technology for degrading Bacillus subtilis and engineering bacteria, which is applied in the field of constructing and applying microbial engineering bacteria, can solve the problems of low enzyme yield, long period of degrading lignin, slow growth of white rot fungi, etc., and achieves rapid growth and reproduction and low nutritional requirements. Effect
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[0022] Example one
[0023] 1.1 Obtain the enzyme gene and the relevant original documents needed to construct the expression vector
[0024] (1) PCR amplification of the glyceraldehyde triphosphate dehydrogenase promoter sequence of Bacillus megaterium.
[0025] Extract the genomic DNA of Bacillus megaterium, use the genomic DNA as a template, and apply primer 5’TT TACGTA GATCCATTATCGGTGAACCA3’ and 5’GG GCATGC GGGAATACATTACGGCCGAT3') was subjected to PCR amplification, and the PCR product was sequenced and analyzed with the BLAST software provided by NCBI to prove that it was the promoter sequence of the glyceraldehyde triphosphate dehydrogenase gene.
[0026] (2) PCR amplification of the laccase gene of Bacillus subtilis
[0027] Use snailase to lyse the cell wall of Bacillus subtilis, extract Bacillus subtilis genomic DNA, and apply the upstream primer (5’AA CCTAGG ATGACACTTGAAAAATTTGTGGATGC3’) and downstream primers (5’AA GCGGCCGC CTATTTATGGGGATCAGTTATA3') was subjected to PCR a...
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[0059] Example two
[0060] 2.1 Obtain the enzyme gene and the relevant original documents needed to construct the expression vector
[0061] (1) PCR amplification of the glyceraldehyde triphosphate dehydrogenase promoter sequence of Bacillus megaterium.
[0062] Extract the genomic DNA of Bacillus megaterium, use the genomic DNA as a template and apply primer 5’TT TACGTA GATCCATTATCGGTGAACCA3’ and 5’GG GCATGC GGGAATACATTACGGCCGAT3') was subjected to PCR amplification, and the PCR product was sequenced and analyzed with the BLAST software provided by NCBI to prove that it was the promoter sequence of the glyceraldehyde triphosphate dehydrogenase gene.
[0063] (2) PCR amplification of the laccase gene of Bacillus subtilis
[0064] Use snailase to lyse the cell wall of Bacillus subtilis, extract Bacillus subtilis genomic DNA, and apply upstream primer (5’AA CCTAGG ATGACACTTGAAAAATTTGTGGATGC3’) and downstream primer (5’AA GCGGCCGC CTATTTATGGGGATCAGTTATA3') was subjected to PCR amplifi...
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