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Immune Protective Efficacy of Recombinant Outer Membrane Protein H of Pasteurella multocida B:2 Associated with Hemorrhagic Septicemia in Pakistan

Immune Protective Efficacy of Recombinant Outer Membrane Protein H of Pasteurella multocida B:2 Associated with Hemorrhagic Septicemia in Pakistan

Abida Mushtaque1, Ali Ahmad Sheikh1, Aamir Ghafoor1*, Wasim Shehzad2 and Nadeem Ahmad3

1Institute of Microbiology, University of Veterinary and Animal Sciences, Outfall Road, Lahore, Pakistan
2Department of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Outfall Road, Lahore, Pakistan
3Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan
 
*      Corresponding author: aamir.ghafoor@uvas.edu.pk

Fig. 1.

Molecular identification of P. multocida B:2 and Omph (A) M: O’Gene Ruler 100bp Plus DNA ladder, lane 1: the 480 bps species conserver region amplified by using KMT1T7 and KMT1SP6 primers, lane 2: negative control, lane 3: positive control; (B) M: O’Gene Ruler 100bp Plus DNA ladder, lane 1: the 620 bps serotype specific region amplified by using KMT1T7 and KMT1SP6 primers, lane 2: positive control, lane 3: negitive control. (C) M: O’Gene Ruler 1 kb DNA ladder, lane 1-10: amplified omph amplificaon, lane 11: positive control, lane 12: negitive control.

Fig. 2.

(A) Restriction digestion of pTZ57R/T plasmid cloned with ompH gene. The enzyme digesation confirmed the presence of gene of interest omph characterized by appearance of 1002 bps band and pTZ57R/T plasmid characterized by 2886 bps band (lane 1 and 2), M: O’Gene Ruler 1 kb DNA ladder. (B) the plasmid (pET-40b(+):omph) was digested with BamHI and SalI to futher confirm the cloning. The presence of two bands of 1002 and 6190 bps, confirmed the successful cloning of omphinpET-40(+) vector.

Fig. 3.

Total cell protein analysis of induced and uninduced transformed BL21 (DE3). (A) M is Page ruler plus prestained protein ladder, Lane 1-3 correspond to the total cell protein of induced BL21 (DE3) transformed cells and lane 4 corresponds to the total cell protein of uninduced BL21 (DE3) cells termed as negative control. (B) Qualitative analysis of purified rOmph protein (Lane 1, 2 and 3). (C) Immunoblotting of rOmpH protein. M: PageRuler protein ladder, lane 1-2: purified rOmph protein, lane 3: negative control, lane 4: histidine tag containing protein used as positive control.

Pakistan Journal of Zoology

February

Pakistan J. Zool., Vol. 56, Iss. 1, pp. 01-501

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