A codon-optimized Chlamydia trachomatis ctl0286 gene and its application

[0027] The codon-optimized ct10286 gene sequence is shown in SEQ ID NO.1.

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[0039] The PCR product was added with 1 μL of DpnI, digested at 37°C for 1 h to remove the template plasmid, and stored at -20°C.

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[0042] 6. The recombinant product is transformed. The above recombinant products were immediately transformed into Escherichia coli DH5α competent cells and spread on the phase

[0049] (7) Invert overnight culture in a 37°C incubator.

[0054] 9. Identification of protein expression. Figure 2 is Coomassie brilliant blue staining to verify the expression of CTL0286 protein. Take the above 1mL culture

[0056] 11. Protein purification. The above-purified inclusions were resuspended in 10 mL of denatured lysate (25 mM HEPES

[0058] Preparation of CTL0286 polyclonal antibody. Replace the NT-CTL0286 protein solution obtained above with PBS buffer

[0059] Figure 3 shows the ability of the CTL0286 polyclonal antibody to recognize the protein. Figure A shows the identification of antibodies from 4 different mice

[0060] The function of the purified protein was verified by protein binding experiments. Figure 4 shows the binding energy of CTL0286 to cRNAPβ' subunit