Over expression of the constitutive expression refractory Mycobacterium tuberculosis Rv3488 protein with an acid-inducible expression system

Pham, Nam Ngoc; Dao, Nelson; Saviola, Beatrice

doi:10.1016/j.mimet.2018.09.021

Cited by 1 publication

(1 citation statement)

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“…Consistent with this fact, our results demonstrated that a low concentration of IPTG (0.3 mmol/L) was advantageous for OmpA expression. Some research studies found that low temperature is also beneficial for protein expression [18,19], which supports our results. Thus, our data suggest that optimal fermentation production conditions involve strain induction at the logarithmic growth period, low IPTG concentration, and low temperature.…”

Section: Discussionsupporting

confidence: 92%

Immunoprotective evaluation of <I>Escherichia coli</I> outer membrane protein A against the main pathogens of animal mastitis

Chen¹,

Wu²,

Na³

et al. 2020

Trop. J. Pharm Res

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Purpose: To evaluate prokaryotic expression of the Escherichia coli (E. coli) outer membrane protein A (OmpA) and its immunoprotective function against the main pathogens of animal mastitis.Methods: A molecular cloning method was used to develop a prokaryotic strain expressing OmpA protein, which was purified by Ni-affinity chromatography. Polyclonal antiserum was generated in mice immunized with OmpA protein. Enzyme-linked immunosorbent assay (ELISA) and western blotting were used to determine the titer and verify anti-OmpA serum specificity, respectively. Interaction between OmpA antiserum and main pathogens of animal mastitis was verified by ELISA and a pull-down method. The immune protective function of OmpA protein was evaluated in mice challenged with pathogens of animal mastitis. Optimal fermentation conditions to produce OmpA protein were determined by the L9(34) orthogonal test.Results: A prokaryotic strain expressing OmpA protein was developed, and purified OmpA was used to develop a mouse polyclonal antibody. The anti-OmpA serum exhibited high specificity and a titer of 1:1600. Anti-OmpA serum directly interacted with E. coli and Staphylococcus aureus (S. aureus). OmpA demonstrated a significant immune protective function of 58.33 % against E. coli and 46.15 % against S. aureus. The optimal conditions for expressing fermentation OmpA were a strain absorbance of 0.5 at a wavelength of 600 nm, IPTG final concentration of 0.3 mmol/L, induction time of 12 h, and induction temperature of 28 °C.Conclusion: OmpA possesses selective immunogenicity and a significant immune protective effect against the main pathogens of animal mastitis. The results suggest that OmpA may potentially be used as a vaccine for animal mastitis. Keywords: E. coli, OmpA protein, Immunoprotection, Animal mastitis, Protein fermentation

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Section: Discussionsupporting

confidence: 92%