Tubercle bacillus fusion protein Mtb8.4-HspX, and preparation method and application thereof
A technology of mtb8.4-hspx and fusion protein, applied in chemical instruments and methods, bacteria, hybrid peptides, etc., can solve unconsidered and affected problems, and achieve the effect of strong cellular immune activity
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[0047] 1. Construction of recombinant plasmids
[0048] The purified PCR products Mtb8.4 and Hspx and plasmid PET30a were digested with restriction endonucleases Nde I and Sac I respectively, and the digested Mtb8.4T and Hspx gene fragments and plasmid PET30a DNA fragments were passed through T4 DNA ligase After connecting overnight at 16°C, transform into Escherichia coli strain DH5α, spread on LB plates containing 50 mg / mL kanamycin, culture overnight at 37°C, pick positive clones, extract plasmids, and perform PCR and sequencing Identification. The recombinant plasmids PET30a-Mtb8.4 and PET30a-Hspx extracted and sequenced correctly were digested with restriction endonucleases Sac I and HindIII respectively, and Hspx and Mtb8.4 were digested with Sac I and HindIII respectively. After gel recovery and purification, the target gene fragments were ligated with the recovered products of PET30a-Mtb8.4 and PET30a-Hspx vectors under the action of T4 DNA ligase at 16°C overnight to...
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