| تعداد نشریات | 282 |
| تعداد نشریات سازمان | 80 |
| تعداد شمارهها | 3,208 |
| تعداد مقالات | 35,560 |
| تعداد مشاهده مقاله | 53,002,498 |
| تعداد دریافت فایل اصل مقاله | 92,287,915 |
مهندسی یک پروتئین کایمریک نوترکیب متشکل از آنتیژن OmpL متصل شده به ادجوانت مولکولی HBHA (HBHA-OMPL) با هدف توسعه واکسن بر علیه سالمونلا تیفی موریوم | ||
| تحقیقات دامپزشکی و فرآوردههای بیولوژیک | ||
| مقاله 12، دوره 36، شماره 2، تیر 1402، صفحه 100-111 اصل مقاله (3.08 M) | ||
| نوع مقاله: مقاله کامل | ||
| شناسه دیجیتال (DOI): 10.22092/vj.2022.359453.1995 | ||
| نویسندگان | ||
| مرضیه بازگیر1؛ نعمت شمس* 2؛ علی فروهرمهر3؛ امین جایدری4؛ نرگس نظیفی2 | ||
| 1گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه لرستان,، ایران | ||
| 2گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه لرستان، ایران | ||
| 3گروه علوم دام، دانشکده کشاورزی، دانشگاه لرستان، ایران | ||
| 4گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه لرستان، ایران | ||
| چکیده | ||
| سالمونلا تیفی موریوم یک باکتری گرم منفی از خانواده آنتروباکتریاسه است که بعنوان عامل ایجاد کننده بیماری در انسان و دام مطرح میباشد. یکی از جدیدترین راه های مقابله با این عامل بیماریزا استفاده از واکسنهای نوترکیب است. در این مطالعه، سازهای متشکل از آنتیژن OmpL از باکتری سالمونلا تیفــیموریــوم و پروتئین HBHA به عنوان ادجوانت مولکولی از باکتری مایکوباکتریوم به روش درون رایانه ای (in silico) و با استفاده از یک لینکر پپتیدی طراحی شد. بدین منظور توالی نوکلئوتیدی هریک از این پروتئینها از پایگاه دادهها استخراج شدند و سازه نوترکیب HBHA-OmpL با بررسی فریم صحیح خوانش طراحی شد. سپس برای ارزیابی امتیاز ایمنیزایی و خواص فیزیکوشیمیایی و همچنین ساختارهای دوم و سوم این سازه از سرورهای معتبر آنلاین استفاده شد. بهترین مدل سه بعدی پالایش شده برای فرایند داکینگ مولکولی مورد استفاده قرار گرفت. توالی نکلئوتیدی سازه کایمریک نوترکیب برای بیان در اشریشیا کولای بهینه سازی گردید و امکان کلون توالی بهینه شده در وکتور بیانی (+)pET22b بررسی شد. بر اساس نتایج بدست آمده در این مطالعه، سازه کایمریک HBHA- OmpL با وزن مولکولی 45/49 کیلو دالتون دارای شاخص ناپایداری 75/32 و شاخص آنتی ژنسیتی 688/0 بود و ساختارهای مارپیچ آلفا و سیم پیچهای تصادفی بیشترین سهم را در توزیع ساختار دوم این سازه داشتند. بررسی داکینگ پروتئین-پروتئین نشان داد که پروتئین HBHA علیرغم کانژوگه شدن با پروتئین OmpL، توانست از طریق 23 پیوند هیدروژنی به گیرنده MD2/TLR4متصل شود. نتایج کلونینگ درون رایانه ای نیز نشان داد سازه کایمریک میتواند در وکتور (+)pET22b با موفقیت کلون شود. | ||
| کلیدواژهها | ||
| واکسن نوترکیب؛ سالمونلا تیفیموریوم؛ OmpL؛ HBHA | ||
| عنوان مقاله [English] | ||
| Engineering a Recombinant Chimeric Protein Consisting of OmpL Antigen Conjugated to HBHA Molecular Adjuvant (HBHA-OmpL) with the Aim of Developing a Vaccine Against Salmonella Typhimurium | ||
| نویسندگان [English] | ||
| Marzieh Bazgir1؛ Nemat Shams2؛ Ali Forouharmehr3؛ Amin Jaydari4؛ Narges Nazifi2 | ||
| 1,,Department of Pathobiology,Faculty of Veterinary Medicine, Lorestan University, Iran | ||
| 2Department of Pathobiology, Faculty of Veterinary Medicine, Lorestan University, Iran | ||
| 3Department of Animal Science, Faculty of Agriculture, Lorestan University, Iran | ||
| 4Department of Pathobiology, Faculty of Veterinary Medicine, Lorestan University, Iran | ||
| چکیده [English] | ||
| Salmonella Typhimurium is a zoonotic gram-negative bacterium of the Enterobacteriaceae family. One of the newest ways against the pathogen is the use of recombinant vaccines. In this study, a recombinant construct consisting of OmpL antigen from Salmonella Typhimurium and HBHA protein as a molecular adjuvant from Mycobacterium was designed by an in silico method and a peptide linker. For this purpose, sequences of these proteins were extracted from the database and the HBHA-OmpL recombinant construct was designed based on the correct reading frame. Then, reliable online servers were used to evaluate the immunogenicity score and physicochemical properties as well as the secondary and tertiary structures of the designed construct. The best refined 3D model was used for the molecular docking process. A codon optimization was done for expression in E. coli, and the possible cloning of the optimized nucleotide sequence in pET22b(+) expression vector was evaluated. Based on the results obtained in this study, the HBHA-OmpL chimeric construct with a molecular weight of 49.45 kDa had an instability index of 32.75 and an antigenicity index of 0.688, and alpha helices and random coil structures had the largest contribution in its secondary structure. The protein-protein docking results showed that despite being conjugated with OmpL protein, HBHA molecule was able to connect to TLR4/MD2 receptor by 23 hydrogen bonds. Also, cloning results confirmed that nucleotide sequence of the designed vaccine can be successfully inserted in pET22b(+) vector. | ||
| کلیدواژهها [English] | ||
| Recombinant vaccine, Salmonella typhimurium, OmpL, HBHA | ||
| مراجع | ||
|
1- Andersen, D. C. and L. Krummen. 2002. Recombinant protein expression for therapeutic applications. Current opinion in biotechnology 13: 117-123. 2- Attwood, T. K., A. Gisel, N.-E. Eriksson and E. Bongcam-Rudloff. 2011. Concepts, historical milestones and the central place of bioinformatics in modern biology: a European perspective. Bioinformatics-trends and methodologies 1. 3- Brenner, F., R. Villar, F. Angulo, R. Tauxe and B. Swaminathan. 2000. Salmonella nomenclature. Journal of clinical microbiology 38: 2465-2467. 4- de Freitas, C. G., Â. P. Santana, P. H. C. da Silva, V. S. P. Gonçalves, M. d. A. F. Barros, F. A. G. Torres, L. S. Murata and S. Perecmanis. 2010. PCR multiplex for detection of Salmonella Enteritidis, Typhi and Typhimurium and occurrence in poultry meat. 139: 15-22. 5- Delany, I., R. Rappuoli and K. L. Seib. 2013. Vaccines, reverse vaccinology, and bacterial pathogenesis. Cold Spring Harbor perspectives in medicine 3: a012476. 6- Eng, S.-K., P. Pusparajah, N.-S. Ab Mutalib, H.-L. Ser, K.-G. Chan and L.-H. Lee. 2015. Salmonella: a review on pathogenesis, epidemiology and antibiotic resistance. Frontiers in Life Science 8: 284-293. 7- Esposito, C., D. Marasco, G. Delogu, E. Pedone and R. Berisio. 2011. Heparin-binding hemagglutinin HBHA from Mycobacterium tuberculosis affects actin polymerisation. Biochemical and biophysical research communications 410: 339-344. 8- Forouharmehr, A. 2021. Engineering an efficient poly-epitope vaccine against Toxoplasma gondii infection: a computational vaccinology study. Microbial Pathogenesis 152: 104646. 9- Forouharmehr, A., N. Nazifi, S. M. Mousavi and A. Jaydari. 2022. Designing an efficient epitope-based vaccine conjugated with a molecular adjuvant against Bovine Babesiosis: A computational study. Process Biochemistry 121: 170-177. 10- Freeman Jr, T. C., S. J. Landry and W. C. Wimley. 2011. The prediction and characterization of YshA, an unknown outer-membrane protein from Salmonella typhimurium. Biochimica et Biophysica Acta (BBA)-Biomembranes 1808: 287-297. 11- Fulop, M., R. Manchee and R. Titball. 1995. Role of lipopolysaccharide and a major outer membrane protein from Francisella tularensis in the induction of immunity against tularemia. Vaccine 13: 1220-1225. 12- Gamage, D. G., A. Gunaratne, G. R. Periyannan and T. G. Russell. 2019. Applicability of instability index for in vitro protein stability prediction. Protein and peptide letters 26: 339-347. 13- Gheibihayat, S. M., A, Sadeghinia. sh, Nazarian. Z, Adeli. 2015. Mechanisms and Performances of Adjuvants in Vaccine Immunogenicity. Journal of Applied Biotechnology Reports. 2 (3) 257-264. 14- Guruprasad, K., B. B. Reddy and M. W. Pandit. 1990. Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Engineering, Design and Selection 4: 155-161. 15-Jamshidi, A. E., M. Basami and N. S. Afshari. 2009. Identification of Salmonella spp. and Salmonella typhimurium by a multiplex PCR-based assay from poultry carcasses in Mashhad-Iran. International Journal of Veterinary Research. 3 (1) 43-48. 16- Jaydari, A., A. Forouharmehr and N. Nazifi. 2019. Determination of immunodominant scaffolds of Com1 and OmpH antigens of Coxiella burnetii. Microbial pathogenesis 126: 298-309. 17- Jaydari, A., N. Nazifi and A. Forouharmehr. 2020. Computational design of a novel multi-epitope vaccine against Coxiella burnetii. Human Immunology 81: 596-605. 18- Kanzler, H., F. J. Barrat, E. M. Hessel and R. L. Coffman. 2007. Therapeutic targeting of innate immunity with Toll-like receptor agonists and antagonists. Nature medicine 13: 552-559. 19- Kardani, K., A. Bolhassani and A. Namvar. 2020. An overview of in silico vaccine design against different pathogens and cancer. Expert Review of Vaccines 19: 699-726. 20- Kisiela, D. I., S. Chattopadhyay, S. J. Libby, J. E. Karlinsey, F. C. Fang, V. Tchesnokova, J. J. Kramer, V. Beskhlebnaya, M. Samadpour and K. Grzymajlo. 2012. Evolution of Salmonella enterica virulence via point mutations in the fimbrial adhesin. PLoS pathogens 8: e1002733. 21- Kundrotas, P. J., Z. Zhu and I. A. Vakser. 2010. GWIDD: genome-wide protein docking database. 38: D513-D517. 22- Mitov, I., V. Denchev and K. Linde. 1992. Humoral and cell-mediated immunity in mice after immunization with live oral vaccines of Salmonella typhimurium: auxotrophic mutants with two attenuating markers. Vaccine 10: 61-66. 23- Mukkur, T., G. McDowell, B. Stocker and A. Lascelles. 1987. Protection against experimental salmonellosis in mice and sheep by immunisation with aromatic-dependent Salmonella typhimurium. Journal of medical microbiology 24: 11-19. 24- Ngan, G. J. Y., L. M. Ng, R. T. Lin and J. W. Teo. 2010. Development of a novel multiplex PCR for the detection and differentiation of Salmonella enterica serovars Typhi and Paratyphi A. Research in Microbiology 161: 243-248. 25- Parra, M., T. Pickett, G. Delogu, V. Dheenadhayalan, A.-S. Debrie, C. Locht and M. J. Brennan. 2004. The mycobacterial heparin-binding hemagglutinin is a protective antigen in the mouse aerosol challenge model of tuberculosis. Infection and Immunity 72: 6799-6805. 26- Porwollik, S., E. Boyd, C. Choy, P. Cheng, L. Florea, E. Proctor and M. McClelland. 2004. Characterization of Salmonella enterica subspecies I genovars by use of microarrays. Journal of bacteriology 186: 5883-5898. 27- Rashidian, E., A. Forouharmehr, N. Nazifi, A. Jaydari and N. Shams. 2021. Computer-aided design of a novel poly-epitope protein in fusion with an adjuvant as a vaccine candidate against leptospirosis. Current Proteomics 18: 113-123. 28- Rashidian, E., Z. S. Gandabeh, A. Forouharmehr, N. Nazifi, N. Shams and A. Jaydari. 2020. Immunoinformatics approach to engineer a potent poly-epitope fusion protein vaccine against Coxiella burnetii. International Journal of Peptide Research and Therapeutics 26: 2191-2201. 29- Shams, N. and A. Forouharmehr. 2021. Assembling the Most Antigenic Peptides of COVID-19 Immunogenic Proteins Along with a Molecular Adjuvant to Develop a Novel Polyepitope Vaccine: a Bioinformatics Investigation. Vaccine Research 8: 36-46. 30- Shams, N., Z. Shakarami Gandabeh, N. Nazifi, A. Forouharmehr, A. Jaydari and E. Rashidian. 2020. Computational design of different epitope-based vaccines against Salmonella typhi. International Journal of Peptide Research and Therapeutics 26: 1527-1539. 31- Singh, Y., A. Saxena, R. Kumar, P. Bhatt and M. Saxena. 2017. Immunogenic Outer membrane Proteins (Omps) of Salmonella: Potential Candidate for sub-unit vaccine. Virology & Immunology 1: 1-6. 32- Tahamtan, A., J. Charostad, S. J. H. Shokouh and M. Barati. 2017. An overview of history, evolution, and manufacturing of various generations of vaccines. Journal of Archives in Military Medicine 5(3):e12315. 33- Tritto, E., F. Mosca and E. De Gregorio. 2009. Mechanism of action of licensed vaccine adjuvants. Vaccine 27: 3331-3334. 34- Verma, S. K., V. Gautam, K. Balakrishna and S. Kumar. 2009. Overexpression, purification, and immunogenicity of recombinant porin proteins of Salmonella enterica Serovar Typhi (S. Typhi). Journal of microbiology and biotechnology 19: 1034-1040. 35- Werle, M. and A. Bernkop-Schnürch. 2006. Strategies to improve plasma half life time of peptide and protein drugs. Amino acids 30: 351-367. 36- Azizpour A. 2018. A Survey on Prevalence of Salmonella enteritidis and Salmonella typhimurium Serotypes in Broiler Flocks of Ardabil Province and Determination of Their Antibiotics Resistance to Five Antibacterial Agents Widely Used in the Iranian Medical Field .J health 9 (2) :143-151. 37- Shapouri, R., M, Rahnama. Sh, Iqbalzadeh. 2018. Investigating the prevalence of Salmonella serotypes in chicken meat and eggs and determining their antibiotic sensitivity in Zanjan city. Physiology and animal development (biological sciences). 2018; 2(3) 63-71 | ||
|
آمار تعداد مشاهده مقاله: 776 تعداد دریافت فایل اصل مقاله: 474 |
||