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Trigger factor assisted soluble expression of recombinant spike protein of porcine epidemic diarrhea virus in Escherichia coli

Overview of attention for article published in BMC Biotechnology, May 2016
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Title
Trigger factor assisted soluble expression of recombinant spike protein of porcine epidemic diarrhea virus in Escherichia coli
Published in
BMC Biotechnology, May 2016
DOI 10.1186/s12896-016-0268-7
Pubmed ID
Authors

Da-Chuan Piao, Do-Woon Shin, In-Seon Kim, Hui-Shan Li, Seo-Ho Oh, Bijay Singh, S. Maharjan, Yoon-Seok Lee, Jin-Duck Bok, Chong-Su Cho, Zhong-Shan Hong, Sang-Kee Kang, Yun-Jaie Choi

Abstract

Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric pathogen of swine. The spike glycoprotein (S) of PEDV is the major immunogenic determinant that plays a pivotal role in the induction of neutralizing antibodies against PEDV, which therefore is an ideal target for the development of subunit vaccine. In an attempt to develop a subunit vaccine for PEDV, we cloned two different fragments of S protein and expressed as glutathione S-transferase (GST)-tagged fusion proteins, namely rGST-COE and rGST-S1D, in E.coli. However, the expression of these recombinant protein antigens using a variety of expression vectors, strains, and induction conditions invariably resulted in inclusion bodies. To achieve the soluble expression of recombinant proteins, several chaperone co-expression systems were tested in this study. We firstly tested various chaperone co-expression systems and found that co-expression of trigger factor (TF) with recombinant proteins at 15 °C was most useful in soluble production of rGST-COE and rGST-S1D compared to GroEL-ES and DnaK-DnaJ-GrpE/GroEL-ES systems. The soluble rGST-COE and rGST-S1D were purified using glutathione Sepharose 4B with a yield of 7.5 mg/l and 5 mg/l, respectively. Purified proteins were detected by western blot using mouse anti-GST mAb and pig anti-PEDV immune sera. In an indirect ELISA, purified proteins showed immune reactivity with pig anti-PEDV immune sera. Finally, immunization of mice with 10 μg of purified proteins elicited highly potent serum IgG and serum neutralizing antibody titers. In this study, soluble production of recombinant spike protein of PEDV, rGST-COE and rGST-S1D, were achieved by using TF chaperone co-expression system. Our results suggest that soluble rGST-COE and rGST-S1D produced by co-expressing chaperones may have the potential to be used as subunit vaccine antigens.

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Mendeley readers

The data shown below were compiled from readership statistics for 24 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Korea, Republic of 1 4%
China 1 4%
Unknown 22 92%

Demographic breakdown

Readers by professional status Count As %
Student > Master 8 33%
Researcher 6 25%
Student > Bachelor 3 13%
Student > Ph. D. Student 1 4%
Other 1 4%
Other 2 8%
Unknown 3 13%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 9 38%
Agricultural and Biological Sciences 5 21%
Chemistry 2 8%
Computer Science 1 4%
Nursing and Health Professions 1 4%
Other 1 4%
Unknown 5 21%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 04 May 2016.
All research outputs
#5,787,608
of 7,651,252 outputs
Outputs from BMC Biotechnology
#354
of 466 outputs
Outputs of similar age
#186,569
of 266,363 outputs
Outputs of similar age from BMC Biotechnology
#17
of 21 outputs
Altmetric has tracked 7,651,252 research outputs across all sources so far. This one is in the 13th percentile – i.e., 13% of other outputs scored the same or lower than it.
So far Altmetric has tracked 466 research outputs from this source. They receive a mean Attention Score of 4.6. This one is in the 8th percentile – i.e., 8% of its peers scored the same or lower than it.
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We're also able to compare this research output to 21 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.