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Cloning and Expression of Truncated Spike (Sf200) Glycoprotein of Infectious Bronchitis Virus (IBV) in Escherichia coli, and its Immunogenicity to Mice

Cloning and Expression of Truncated Spike (Sf200) Glycoprotein of Infectious Bronchitis Virus (IBV) in Escherichia coli, and its Immunogenicity to Mice

Basit Zeshan1,2,*, Mushtaq A. Saleem2,Javed Iqbal Wattoo2, Mohd Mokhtar Arshad1 and Maizan Mohamed1

1Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kota Bharu 16100, Kelantan, Malaysia
2Faculty of Life Sciences, University of Central Punjab, Lahore, Pakistan

*      Corresponding author: dr.basit@ucp.edu.pk

 

 

Fig. 1.

Gel electrophoresis showing restriction enzyme digestion of complete S1 gene (1650bp approx.) after digestion with XhoI while 1kb DNA ladder is on the other side.

Fig. 2.

Amplification of three fragments of S1 with the size of 150bp, 162bp and 351bp, respectively. Arrow indicate 1kb DNA ladder.

Fig. 3.

Restriction enzyme analysis of pET-32a-Sf200 after digestion with BamHI and XhoI (Lane 1, 3). 1kb DNA ladder is on the other side.

Fig. 4.

 SDS-PAGE of the Purified Sf200 Lane (1) and the cell lysate of E. coli harboring the recombinant plasmid pET-32a-Sf200 (38kDa approx.) after induction with IPTG containing E. coli BL21 (Lane 4), E. coli Rosetta strain (Lane 5), before induction with IPTG (Lane 2), pET-32a (+) vector control (Lane 3) while the proteins standards are showed on right side of panel.

Fig. 5.

Western blot analysis prokaryotic expressed Sf200 in BL21 E. coli (Lane 2), while the proteins standards are showed on left side of panel.

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Pakistan Journal of Zoology

June

Vol. 50, Iss. 3, Pages 799-1198

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