Cloning and expression of recombinant O-acetylserine lyase from human streptococcus suis serotype 2 in pichia pastoris
Article Sidebar
Main Article Content
Abstract
Streptococcus suis is recognized worldwide as an important swine pathogen, which occasionally infects humans and causes fatal illnesses. O-acetylserine lyase (OASL) is responsible for the fi nal step of the cysteine biosynthesis, a key step in bacterial sulfur metabolism and offers a means by which inorganic sulfur maybe incorporated into cellular components. Recently, OASL was reported as a putative virulence-associated factor and immunogenic protein in S. suis. And this suggests that it may be involved in pathogenesis. In this study, the gene coding for OASL was cloned from a clinical S. suis serotype 2 strain. A full-length gene encoding OASL was amplifi ed by the OASL gene-specifi c primer. The amplifi ed fragments were cloned into pGAPZA vector and expression as a His-tagged recombinant protein in Pichia pastoris. The expressed sample collected from the supernatant was fi rst analyzed by SDS-PAGE. After that the recombinant proteins were detected by Ni2+-NTA enzyme conjugate. The Western immunoblotting showed that either antibodies present in patient serum suffered from S. suis infection or rabbit serum raised against whole cell lysate of homologous and heterologous strains can bind to recombinant OASL. These results verify that OASL had antigenicity and guide for evaluating its antigenic potential in further studies.
Article Details
Publishing an article with open access in Veterinary Integrative Sciences leaves the copyright with the author. The article is published under the Creative Commons Attribution License 4.0 (CC-BY 4.0), which allows users to read, copy, distribute and make derivative works from the material, as long as the author of the original work is cited.
References
Arends, J. P., Zanen, H. C. (1988). Meningitis caused by Streptococcus suis in humans. Rev Infect Dis, 10(1), 131-137.
Fernández M, K. M., Kuipers OP, Siezen RJ, van Kranenburg R. (2002). Regulation of the metC-cysK operon, involved in sulfur methabolism in Lactococcus lactis. Journal of Bacteriology, 184(1), 82-90.
Fongcom, A., Pruksakorn, S., Mongkol, R., Tharavichitkul, P., & Yoonim, N. (2001). Streptococcus suis infection in northern Thailand. Journal of the Medical Association of Thailand, 84(10), 1502 1508.
Gottschalk, M., & Segura, M. (2000). The pathogenesis of the meningitis caused by Streptococcus suis: the unresolved questions. Veterinary Microbiology, 76(3), 259-272.
Hell, R. (1997). Molecular physiology of plant sulfur metabolism. Planta, 202(2), 138–148. Hill CE, M. D., MacInnes JI. (2003). A search for virulence genes of Haemophilus parasuis using differential display RT-PCR. Veterinary Microbiology 17, 189-202.
Jeff D. Evans, S. F. A.-K., Scott A. Martin. (2002). Cloning of the O-Acetylserine Lyase Gene from the Ruminal Bacterium Selenomonas ruminantium HD4. CURRENT MICROBIOLOGY, 44, 161–166.
Joseph Sambrook, D. W. R. (2001). Molecular Cloning: A Laboratory Manual (3 ed. Vol. 1). New York: Cold Spring Harbor Laboratory Press. Kanruethai Wongsawan, A. W., Khemika Lomthaisong, Anuson Boonthum, Volaluck Supjatura, and Prasit Thraravichitkul. (2010). Immunoproteomic Assay of Streptococcus suis Serotype 2 Isolated from Patients. Chiang Mai Journal of Science., 37(1), 134-150.
Leelarasamee, A., Nilakul, C., Tien-Grim, S., Srifuengfung, S., & Susaengrat, W. (1997). Streptococcus suis toxic-shock syndrome and meningitis. Journal of the Medical Association of Thailand, 80(1), 63-68.
Lithgow JK, H. E., Cohen G, Aharonowitz Y, Foster SJ. (2004). Role of a cysteine synthase in Staphylococcus aureus. Journal of Bacteriology, 186(6), 1579-1590.
Lun, Z. R., Wang, Q. P., Chen, X. G., Li, A. X., & Zhu, X. Q. (2007). Streptococcus suis: an emerging zoonotic pathogen. The Lancet Infectious Diseases, 7(3), 201-209.
Osaki, M., Takamatsu, D., Tsuji, N., & Sekizaki, T. (2000). Cloning and characterization of the gene encoding O-acetylserine lyase from Streptococcus suis. Current Micro biology, 40(1), 67-71.
Staats, J. J., Feder, I., Okwumabua, O., & Chengappa, M. M. (1997). Streptococcus suis: past and present. Veterinary Research Communications, 21(6), 381-407.
Stancik LM, S. D., Schmidt B, Barnhart DM, Yoncheva YN, Slonczewski JL. ( 2002 ). pH-dependent expression of periplasmic proteins and amono acid catabolism in Escherichia coli. Journal of Bacteriology, 184(15), 4246-4258.
Takeuchi D, K. A., Pienpringam A, Loetthong P, Samerchea S, Luangsuk P, Khamisara K, Wongwan N, Areeratana P, Chiranairadul P, Lertchayanti S, Petcharat S, Yowang A, Chaiwongsaen P, Nakayama T, Akeda Y, Hamada S, Sawanpanyalert P, Dejsirilert S, Oishi K. (2012). Population-based study of Streptococcus suis infection in humans in Phayao Province in northern Thailand. PLoS One, 7(2), e31265-e31265.
Walker J, V. J., Gomez MA, Drummelsmith J, Burchmore R, Girard I, Ouellette M. (2006). Identifi cation of developmentally regulated proteins in Leishmania pana mensis byproteome profi ling of promastigotes and axenic amastigotes. Molecular and Biochemical Parasitology, 147(1), 64-73.
Wangkaew, S., Chaiwarith, R., Tharavichitkul, P., & Supparatpinyo, K. (2006). Streptococcus suis infection: a series of 41 cases from Chiang Mai University Hospital. Journal of Infection, 52(6), 455-460.
Wangsomboonsiri, W., Luksananun, T., Saksornchai, S., Ketwong, K., & Sungkanuparph, S. (2008). Streptococcus suis infection and risk factors for mortality. Journal of Infection, 57(5), 392-396.
Wu, Z., Zhang, W., & Lu, C. (2008). Comparative proteome analysis of secreted proteins of Streptococcus suis serotype 9 iso lates from diseased and healthy pigs. Microbial Pathogenesis, 45(3), 159-166.
Zhang, W., & Lu, C. P. (2007). Immunoproteomic assay of membrane-associated proteins of Streptococcus suis type 2 China vaccine strain HA9801. Zoonoses Public Health, 54(6-7), 253-259.