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Treponema pallidum recombinant chimeric antigen and its preparation method and use

A technique of Treponema pallidum, chimeric antigen

Active Publication Date: 2021-12-03
SICHUAN ANKERUI NEW MATERIAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these recombinant antigens have more or less certain defects in the preparation process or in the process of immunological detection, which limits the application of recombinant chimeric antigens in the detection of Treponema pallidum

Method used

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  • Treponema pallidum recombinant chimeric antigen and its preparation method and use
  • Treponema pallidum recombinant chimeric antigen and its preparation method and use
  • Treponema pallidum recombinant chimeric antigen and its preparation method and use

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 present invention relates to a recombinant chimeric antigen of Treponema pallidum, which consists of the amino acid sequence shown in SEQ ID NO.12, the first flexible linker, the amino acid sequence shown in SEQ ID NO.9, the second flexible linker It is formed in series with the amino acid sequence shown in SEQ ID NO.13. The recombinant chimeric antigen of the invention can be used for serological detection of treponema pallidum antibody, and has improved dissolution expression ability, activity and thermal stability. The present invention also relates to a method for preparing the recombinant chimeric antigen.

Description

technical field [0001] The invention relates to the field of immunological detection, in particular to a Treponema pallidum recombinant chimeric antigen. Background technique [0002] Syphilis is a chronic, systemic sexually transmitted disease caused by Treponema pallidum (TP; also known as Treponema pallidum). In the "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases", syphilis is listed as a category B disease for prevention and management . Its route of transmission is mainly sexual transmission, clinically syphilis is divided into primary syphilis, secondary syphilis, tertiary syphilis, latent syphilis and congenital syphilis (congenital syphilis) and so on. [0003] Syphilis is an infectious disease that is prevalent all over the world. According to WHO estimates, there are about 12 million new cases worldwide every year, mainly concentrated in South Asia, Southeast Asia and sub-Saharan Africa. In recent years, syphilis has gr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/70C07K19/00C12N15/62G01N33/68G01N33/569
CPCC07K14/20C07K2319/00C12N15/70G01N33/56911G01N33/6854G01N2333/20
Inventor 干盈盈韩新鹏吕志强罗琴文丹华
Owner SICHUAN ANKERUI NEW MATERIAL TECH CO LTD
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