Recombinant chimeric antigen of treponema pallidum as well as preparation method and use thereof

A technology of Treponema pallidum and chimeric antigen, which is applied in the field of immunological detection, can solve the problems of restricting the application of recombinant chimeric antigen, and achieve the effect of improving the degree of soluble expression, high activity, high stability and activity

Active Publication Date: 2020-08-25
SICHUAN ANKERUI NEW MATERIAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The technical effect of this patented method improves both the amount of protein produced by an individual's own cells (which increases their ability to fight off disease) while also increasing its overall level of stableness against environmental factors such as heat or pH changes over time. This results in longer-lasting immune responses when used alone without any further treatment.

Problems solved by technology

The technical problem addressed in this patents relates to developing improved ways to detect treponma pneumonia without causing confusion among different diseased individuals due to its high variability between individual populations.

Method used

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  • Recombinant chimeric antigen of treponema pallidum as well as preparation method and use thereof
  • Recombinant chimeric antigen of treponema pallidum as well as preparation method and use thereof
  • Recombinant chimeric antigen of treponema pallidum as well as preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Sequence Design of Wild Type Treponema pallidum Recombinant Chimeric Antigen WTP17+15+47

[0071] Select the full-length sequence of the wild-type TP17 antigen after removing the signal peptide, that is, the 25-156aa region, its amino acid sequence is shown in SEQ ID NO.1, and the corresponding codon-optimized nucleotide sequence is shown in SEQ ID NO.15 shown; select the full-length sequence of the wild-type TP15 antigen after removing the signal peptide, that is, the 17-158aa region, its amino acid sequence is shown in SEQ ID NO.2, and the corresponding codon-optimized nucleotide sequence is shown in SEQ ID NO. 16; select the full-length sequence of the wild-type TP47 antigen after removing the signal peptide, that is, the 20-434aa region, its amino acid sequence is shown in SEQ ID NO.3, and the corresponding codon-optimized nucleotide sequence is shown in SEQ ID Shown in NO.17.

[0072] The above three epitopes are passed through linker 1 (amino acid seque...

Embodiment 2

[0074] Example 2 Construction of recombinant wild-type chimeric antigen 32-WTP17+15+47 and confirmation of its expression form

[0075] Using wtp17+15+47 in Example 1 as a template, using TP17-F (SEQ ID NO.29) and TP47-R (SEQ ID NO.30) as a primer pair, carry out PCR amplification according to the method described in the molecular cloning experiment guide The nucleotide sequence of wtp17+15+47 was obtained by amplification and gel recovery, and the wtp17+15+47 gene fragment was constructed into the pET32a vector, and the resulting recombinant vector was named pET32a-wtp17+15+47.

[0076] Transform pET32a-wtp17+15+47 into Escherichia coli E.coli BL21 (Rossett) competent cells, and induce expression according to the method described in the molecular cloning experiment guide. The brief description is: 37°C, 200rpm culture to OD600=0.8, add IPTG to a final concentration of 1mM, and induce at 25°C for 16 hours. The clone that induced the target protein was named R pET32a-WTP17+15+...

Embodiment 3

[0079] Example 3 Construction of recombinant wild-type chimeric antigen CN-WTP17+15+47 and confirmation of its expression form

[0080] The nucleotide sequence of the SUMO fusion tag (SEQ ID NO.21, wherein the amino acid sequence of the SUMO fusion tag is shown in SEQ ID NO.7) was obtained by chemical synthesis, and used as a template to SUMO-F (SEQ ID NO.31) and SUMO-R (SEQ ID NO.32) were used as a primer pair, and the nucleotide sequence of the SUMO fusion tag was obtained by performing PCR amplification and gel recovery according to the method described in the Molecular Cloning Experiment Guide. Using wtp17+15+47 in Example 1 as a template, using SUMO-TP17-F (SEQ ID NO.33) and TP47-R (SEQ ID NO.30) as a primer pair, according to the method described in the molecular cloning experiment guide Perform PCR amplification and gel recovery to obtain wtp17+15+47 nucleotide sequence. Subsequently, using the nucleotide sequence of wtp17+15+47 and SUMO fusion tag as a template, using...

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Abstract

The invention relates to a recombinant chimeric antigen of treponema pallidum. The recombinant chimeric antigen is formed by connecting an amino acid sequence as shown in SEQ ID NO.12, a first flexible linker, an amino acid sequence as shown in SEQ ID NO.9, a second flexible linker and an amino acid sequence as shown in SEQ ID NO.13 in series. The recombinant chimeric antigen can be used for serological detection of treponema pallidum antibody, and has improved dissolving expression capability, activity and thermal stability. The invention also relates to a method for preparing the recombinantchimeric antigen.

Description

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Claims

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Application Information

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Owner SICHUAN ANKERUI NEW MATERIAL TECH CO LTD
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