MAP of F1 and V antigens from Yersinia pestis astride innate and adaptive immune response

Rai, Reeta; Das, Baijnath; Choudhary, Nageshwar; Talukdar, Ayantika; Rao, Dandina N.

doi:10.1016/j.micpath.2015.07.012

Cited by 2 publications

(2 citation statements)

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“…5 Coomassie brilliant blue staining of protein, expressed in different temperatures, IPTG concentrations and culture medium by Taguchi statistical method. Time zero before induction (1,9), test level one (2), two (3), three (4), protein size marker (5), test level four (6), five (7), six (8), seven (10), eight (11) and nine (12) temperature, IPTG concentration, and medium on the V antigen expression. After optimizing the production conditions, the cells were incubated in the cleavage buffer after incubation at 4 °C for 16 h to obtain a protein with a high purity of 90% and a suitable efficiency, as shown in Fig.…”

Section: Purification Of V Antigenmentioning

confidence: 99%

“…LcrV is known as the virulence and multifunctional protein. This crucial protein has been shown to act at the level of secretion control by binding to other proteins in order to modulate the host immune response by altering cytokine production [7,8]. Genetic engineering can be used to produce recombinant vaccines using different parts of Yersinia, such as V antigen and F1 [9,10].…”

Section: Introductionmentioning

confidence: 99%

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Optimization and One-Step Purification of Recombinant V Antigen Production from Yersinia pestis

et al. 2020

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The purpose of this study was to develop an efficient and inexpensive method for the useful production of recombinant protein V antigen, an important virulence factor for Yersinia pestis. To this end, the synthetic gene encoding the V antigen was subcloned into the downstream of the intein (INT) and chitin-binding domain (CBD) from the pTXB1 vector using specific primers. In the following, the produced new plasmid, pTX-V, was transformed into E. coli ER 2566 strain, and the expression accuracy was confirmed using electrophoresis and Western blotting. In addition, the effects of medium, inducer, and temperature on the enhancement of protein production were studied using the Taguchi method. Finally, the V antigen was purified by a chitin affinity column using INT and CBD tag. The expression was induced by 0.05 mM IPTG at 25 °C under optimal conditions including TB medium. It was observed that the expression of the V-INT-CBD fusion protein was successfully increased to more than 40% of the total protein. The purity of V antigen was as high as 90%. This result indicates that V antigen can be produced at low cost and subjected to one-step purification using a self-cleaving INT tag.

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