Applications of lactobacillin plnE
A technology of lactic acid bacteria and amino acids, applied in the field of genetic engineering, can solve the problems of limited production of lactic acid bacteria, large amount required for antibacterial, low activity, etc., to improve intestinal mucosal immunity, small molecular weight, and convenient artificial synthesis Effect
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[0019] Embodiment: the preparation of lactobacillus plnE
[0020] (1) Refer to the nucleotide sequence of the plnE gene of Gene Bank lactic acid bacteria (Gene Bank: X94434), use Primer Premier6 to design a pair of specific primers, plnE upstream primer F: 5'-CGC GGATCC ATGCTACAGTTTGAGAAGTT-3', plnE downstream primer R: 5'-CG GAATTC TTAACGAATACTTTTCAAAA-3', the designed primer sequence was sent to Shanghai Sangon Bioengineering Co., Ltd. for synthesis.
[0021] (2) The upstream primer F and downstream primer R of the plnE gene were used as amplification primers, and the lactic acid bacteria DNA was used as a template to perform PCR amplification on the plnE gene, and the PCR amplification products were identified by agarose gel electrophoresis.
[0022] (3) After identifying the PCR product by agarose gel electrophoresis, purify the target band by tangent; carry out double digestion with the purified plnE product and plasmid pET-32(+); take the digested PCR target fragment a...
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