Bacillus subtilis neutral phytase phyC gene
A technology of Bacillus subtilis and phytase, applied in plant genetic improvement, genetic engineering, hydrolytic enzymes, etc., can solve the problems of low enzyme activity and limited application range of acid phytase
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[0057] Example: First, primers were designed according to the neutral phytase phyC gene reported on NCBI, the upstream is: 5'TACgTA ATg AAT CAT TCA AAA ACA CTT 3', the downstream is: 5'gCA CTA gTT TAT TTT CCg CTTCTg TCA; Add the SnaBI restriction enzyme site in the upstream; expand the culture of Bacillus subtilis and extract its genome, and then use it as a template to amplify with the designed specific primers to obtain the target band. After the PCR product is recovered, it is combined with pGEM- The T vector was ligated and transformed into Escherichia coli DH5α competent cells, and the positive clones were obtained and sent to Shanghai Invitrogen Company for sequencing. The sequencing results showed that the amplified sequence was consistent with the expected results; the T clone containing the target fragment and the pPIC9K empty plasmid were respectively used with SnaBI Carry out double enzyme digestion with NotI, recover the target fragments respectively, connect them w...
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