Expression, purification and immunochemical characterization of recombinant OMP28 protein of &lt;i&gt;Brucella&lt;/i&gt; species

Manat, Yesbol; Shustov, Alexandr V.; Evtehova, E; Eskendirova, Saule

doi:10.4314/ovj.v6i2.1

Cited by 16 publications

(14 citation statements)

References 18 publications

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“…2.2), which further confirmed the antigenicity of the Omp28 protein. Earlier some researchers reported the formation of inclusion bodies in Omps expression (MANAT et al, 2016;YOUSEFI et al, 2016), but in our study we did not observe any inclusion body formation, even at a higher inducer concentration (IPTG). Omp28 protein is formed by three identical subunits which are not linked by disulfide bonds, representing a molecular mass of 28 kDa (NEVES-FERREIRA et al, 2004).…”

Section: Resultscontrasting

confidence: 83%

Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development - a short communication

Pandey¹,

Saxena²,

Saxena³

et al. 2018

Vet. arhiv

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KUMAR: Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development -a short communication. Vet. arhiv 88, 559-568, 2018.ABSTRACT Salmonella Typhimurium, a major gastrointestinal pathogen, poses a global threat to human health. Public health problems associated with this organism have increased to the extent that it has become a major issue. The bacterium is becoming resistant to the commonly available antibiotics, and vaccines also suffer from limitations such as short lived immunity. Therefore, there is an urgent need for the development of an effective vaccine. The outer membrane proteins (Omps) of Salmonella have proven their capability to be developed as a vaccine candidate for prevention of salmonellosis. With this aim, in the present study the Omp28 gene of Salmonella Typhimurium was amplified, cloned and expressed under an IPTG induction system. The recombinant protein thus produced was purified and tested for its antigenicity. The antigenicity of the purified protein was confirmed by western blotting with antiserum raised in rabbit against Omps of S. Typhimurium. The Omp28 gene was amplified as a 330bp product. The expressed protein was found to be of approximately 28kDa and it produced a strong signal in western blot analysis. This study concluded that Omp28 may be proven to be an effective candidate for the development of r-DNA vaccine against salmonellosis.

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Section: Resultscontrasting

confidence: 83%

Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development - a short communication

Pandey¹,

Saxena²,

Saxena³

et al. 2018

Vet. arhiv

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“…Escherichia coli BL21 strains, producing Brucella recombinant proteins, were obtained as described in our previous studies: B. abortus rOMP25 and B. melitensis rOMP31 [16], rBP26 [14], and rSOD [17].…”

Section: Microbial Culturesmentioning

confidence: 99%

“…Among the known rOMPs, the most studied ones are rOMP25 [12] and rOMP31 [13]. Another group of recombinant proteins, which is of great interest from a diagnostic point of view, are periplasmic proteins: rBP26 or rOMP28 [14] and Cu-Zn superoxide dismutases (rSOD) [15]. However, the comparative diagnostic value of these two groups of Brucella proteins is still scarcely explored.…”

Section: Introductionmentioning

confidence: 99%

Serodiagnostic potential of Brucella outer membrane and periplasmic proteins

Bulashev¹,

Akibekov²,

Suranshiyev³

et al. 2019

Turk J Vet Anim Sci

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The aim of this study was to evaluate the serological diagnostic potential of the Brucella recombinant outer membrane (rOMP25, rOMP31) and periplasmic proteins (rBP26, rSOD) in a comparative way using an indirect enzyme-linked immunosorbent assay (i-ELISA). Rabbit and/or mouse antibodies to Brucella whole cell and/or soluble protein preparations recognized all recombinant proteins used, which confirms the expression of target antigens in E. coli in active form. The recombinant proteins showed different antigenicity to antibodies of cattle kept on a brucellosis-affected (endemic) farm and/or a new focus of infection. Thus, the presence of anti-Brucella antibodies was confirmed by i-ELISA/rSOD in 79% of cows from endemic conditions with positive results by conventional serological tests (RBPT and/or CFT). However, antibodies specific to this protein were detected in only 14% of seropositive animals kept in the hotbed of a new brucellosis infection. Moreover, rSOD-specific antibodies were not detected in the sera of vaccinated cattle from a brucellosis-free farm, whereas antibodies to other recombinant proteins were found in 2%-8% of animals. Using recombinant proteins in immunoassays significantly reduced the number of cows positive for brucellosis. Furthermore, there was not a single protein among the rOMPs that would show the total positive results of all proteins used. Thus, the development of reliable ELISA tests for the diagnosis of brucellosis requires further comprehensive study of the recombinant proteins in order to design a multiprotein antigen that consists of a combination of several proteins with diagnostic potential.

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“…In another study by Manat et al, (2016) a purified recombinant outer membrane protein 28 (rOMP28) of Brucella species produced in Escherichia coli (E. coli) was evaluated as a diagnostic antigen in an Indirect ELISA (I-ELISA) for bovine Brucellosis. The results showed that the rOMP28 of Brucella spp.…”

Section: Biomarkers Of Brucella and Brucellosismentioning

confidence: 99%

Brucellosis in Sheep and Goats and its Serodiagnosis and Epidemiology

Saxena¹,

Saxena²

2018

Int.J.Curr.Microbiol.App.Sci

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Journal homepage: http://www.ijcmas.com Brucellosis is an important zoonotic disease globally that causes huge economic losses to the livestock owners and is of public health significance. Brucellosis in animals is endemic in India. In sheep and goats, Brucellosis is mainly caused by Brucella melitensis whereas Brucella ovis causes the disease in sheep. The symptoms in infected sheep and goats are abortions, stillbirths and the birth of weak offsprings. Animals that abort may retain the placenta. Sheep and goats usually abort only once, but reinvasion of the uterus and shedding of organisms can occur during subsequent pregnancies. Several studies have been carried out on seroepidemiology of caprine and ovine Brucellosis in India. The tests commonly used for diagnosis of Brucellosis are the milk ring test, Rose Bengal Plate Test (RBPT), Standard Tube Agglutination Test (STAT), Microtiter Plate Agglutination Test (MAT) and ELISA. The RBPT is a rapid screening test for the diagnosis of Brucellosis. The sensitivity of RBPT is very high (>99%) but the specificity can be low and it could sometimes give a false positive result. Its positive predictive value is low and a positive test result requires confirmation by a more specific test. Isolation and culture of Brucella organisms is the gold standard test for the diagnosis of Brucellosis. K e y w o r d sOvine Brucellosis, Caprine Brucellosis, Serodiagnosis, Seroepidemiology

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Expression, purification and immunochemical characterization of recombinant OMP28 protein of <i>Brucella</i> species

Cited by 16 publications

References 18 publications

Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development - a short communication

Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development - a short communication

Serodiagnostic potential of Brucella outer membrane and periplasmic proteins

Brucellosis in Sheep and Goats and its Serodiagnosis and Epidemiology

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