Recombinant expression and immunological characterization of hemolysin coregulated protein 1, <italic style="font-style: italic">Burkholderia pseudomallei</italic> type Ⅵ secretion system protein

Meijuan ZHANG; Zhiqiang HU; Yupei XIA; Siqi YUAN; Jingmin YAN; Chenglong RAO; Qian LI; Wenbo YANG; Xuhu MAO

doi:10.16016/j.1000-5404.202008056

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Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein

Basic Medicine | 更新时间：2026-01-09

- Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein
- Journal of Third Military Medical University Vol. 43, Issue 23, Pages: 2296-2301(2020)
- 作者机构：
  
  400038 重庆, 陆军军医大学(第三军医大学)药学与检验医学系临床微生物及免疫学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81971907);the General Program of National Natural Science Foundation of China(31970135)
- DOI：10.16016/j.1000-5404.202008056
  CLC： R378.99;R394-33;R446.62
- Received：07 August 2020，
  
  Revised：2020-09-25，
  
  Published：15 December 2020
- 稿件说明：
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Meijuan ZHANG, Zhiqiang HU, Yupei XIA, et al. Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein[J]. Journal of Third Military Medical University, 2020, 43(23): 2296-2301.
DOI：

Meijuan ZHANG, Zhiqiang HU, Yupei XIA, et al. Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein[J]. Journal of Third Military Medical University, 2020, 43(23): 2296-2301. DOI： 10.16016/j.1000-5404.202008056.

摘要

目的

重组表达类鼻疽杆菌Ⅵ型分泌系统溶血素共调节蛋白1(hemolysin coregulated protein 1，Hcp1)，并对其免疫学性质进行鉴定。

方法

利用DNA重组技术以pET-28a表达系统在

E.coli

BL21(DE3)中重组表达Hcp1；通过亲和层析制备高纯度Hcp1蛋白；采用腹腔注射免疫小鼠制备抗Hcp1血清，ELISA检测抗体滴度，免疫印迹及免疫荧光分析抗体的免疫反应性。

结果

成功获得纯度达95%的重组Hcp1蛋白，该蛋白免疫小鼠后获得的抗血清具有较好的特异性和较高的抗体滴度(1:512 000)，表现出良好的免疫原性和免疫反应性。

结论

成功制备了具有生物学活性的Hcp1重组蛋白及其抗血清。

Abstract

Objective

To express recombinant hemolysin coregulated protein 1 (Hcp1) of type Ⅵ secretion system (T6SS) of

Burkholderia pseudomallei

and identify its immunological properties.

Methods

The recombinant Hcp1 protein was expressed by using DNA recombination technology through pET-28a expression system in

E.coli

BL21(DE3)

and then purified by affinity chromatography. Then 6-week-old BALB/c mice were immunized with the obtained Hcp1

via

intraperitoneal injection

the serum samples were collected to detect the titers of anti-Hcp1 specific antibodies by ELISA. Meanwhile

the immune response against antigen-specific antibodies was evaluated by Western blotting and immunofluorescence assay.

Results

The recombinant Hcp1 protein was successfully obtained

with a purity of 95%. The obtained recombinant protein stimulated the antisera of good specificity and high titer (1: 512 000)

with good immunogenicity and immunoreactivity.

Conclusion

The recombinant protein Hcp1 and its antisera are successfully prepared.

关键词

Keywords

references

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Related Institution

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Department of Clinical Microbiology and Immunology, College of Medical Laboratory Sciences, Southwest Hospital, Third Military Medical University

Department of Clinical Microbiology and Immunology, Faculty of Medical Laboratory Science, Southwest Hospital, Third Military Medical University

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Recombinant expression and immunological characterization of hemolysin coregulated protein 1, Burkholderia pseudomallei type Ⅵ secretion system protein

DOI：10.16016/j.1000-5404.202008056

摘要

Abstract

关键词

Keywords

references