Molecular Cloning and Expression of Pro J 1: A New Allergen of Prosopis Juliflora Pollen
-
Fatemeh Dousti
,
Mohammad-Ali Assarehzadegan
,
Payam Morakabati
,
Gholam Reza Khosravi
,
Bahareh Akbari
Abstract
Pollen from mesquite (Prosopis juliflora) is one of the important causes of immediate hypersensitivity reactions in the arid and semi-arid regions of the world. The aim of present study is to produce and purify the recombinant form of allergenic Ole e 1-like protein from the pollen of this allergenic tree. Immunological and cross-inhibition assays were performed for the evaluation of IgE-binding capacity of purified recombinant protein. For molecular cloning, the coding sequence of the mesquite Ole e 1-like protein was inserted into pTZ57R/T vector and expressed in Escherichia coli using the vector pET-21b(+). After purification of the recombinant protein, its immunoreactivity was analysed by in vitro assays using sera from twenty one patients with an allergy to mesquite pollen. The purified recombinant allergen was a member of Ole e 1-like protein family and consisted of 150 amino acid residues, with a predicted molecular mass of 16.5 kDa and a calculated isoelectric point (pI) of 4.75. Twelve patients (57.14%) had significant specific IgE levels for this recombinant allergen. Immunodetection and inhibition assays indicated that the purified recombinant allergen might be the same as that in the crude extract. Herein, we introduce an important new allergen from P. juliflora pollen (Pro j 1), which is a member of the Ole e 1-like protein family and exhibits significant identity and similarity to other allergenic members of this family.
1. Al-Frayh, A, Hasnain, SM, Gad-El-Rab, MO, Al-Turki, T, Al-Mobeireek, K and Al-Sedairy, ST. Human sensitization to Prosopis juliflora antigen in Saudi Arabia. Ann Saudi Med. 1999;19(4):331-6.
2. Assarehzadegan, M-A, Shakurnia, A and Amini, A. The most common aeroallergens in a tropical region in Southwestern Iran. World Allergy Organ J. 2013;6:7.
3. Assarehzadegan, M-A, Shakurnia, AH and Amini, A. Sensitization to common aeroallergens among asthmatic patients in a tropical region affected by dust storm. J Med Sci. 2013;13(7):592–7.
4. Bener, A, Safa, W, Abdulhalik, S and Lestringant, GG. An analysis of skin prick test reactions in asthmatics in a hot climate and desert environment. Allerg Immunol (Paris). 2002;34(8):281-6.
5. Killian, S and McMichael, J. The human allergens of mesquite (Prosopis juliflora). Clin Mol Allergy. 2004;2(1):8.
6. Thakur, IS. Purification and characterization of the glycoprotein allergen from Prosopis juliflora pollen. Biochem Int. 1991;23(3):449-59.
7. Buys, PJC. Aeroallergens in Namibia: review article. Curr Allergy Clin Immunol. 2013;26(4):210-2.
8. Assarehzadegan, M-A, Khodadadi, A, Amini, A, Shakurnia, A-H, Marashi, SS, Ali-Sadeghi, H, et al. Immunochemical characterization of Prosopis juliflora pollen allergens and evaluation of cross-reactivity pattern with the most allergenic pollens in tropical areas. Iran J Allergy, Asthma Immunol. 2014;14(1):74-82.
9. Dhyani, A, Arora, N, Gaur, SN, Jain, VK, Sridhara, S and Singh, BP. Analysis of IgE binding proteins of mesquite (Prosopis juliflora) pollen and cross-reactivity with predominant tree pollens. Immunobiology. 2006;211(9):733-40.
10. Dhyani, A, Singh, BP, Arora, N, Jain, VK and Sridhara, S. A clinically relevant major cross-reactive allergen from mesquite tree pollen. Eur J Clin Invest. 2008;38(10):774-81.
11. Hasnain, SM, Al-Frayh, AR, Subiza, JL, Fernandez-Caldas, E, Casanovas, M, Geith, T, et al. Sensitization to indigenous pollen and molds and other outdoor and indoor allergens in allergic patients from saudi arabia, United arab emirates, and Sudan. World Allergy Organ J. 2012;5(6):59-65.
12. Suliaman, FA, Holmes, WF, Kwick, S, Khouri, F and Ratard, R. Pattern of immediate type hypersensitivity reactions in the Eastern Province, Saudi Arabia. Ann Allergy Asthma Immunol. 1997;78(4):415-8.
13. Shamsbiranvand, MH, Khodadadi, A, Assarehzadegan, M-A, Borci, SH and Amini, A. Immunochemical characterization of acacia pollen allergens and evaluation of cross-reactivity pattern with the common allergenic pollens. J Allergy (Cairo). 2014;2014:409056.
14. de Dios Alche, J, Mrani-Alaoui, M, Castro, AJ and Rodriguez-Garcia, MI. Ole e 1, the major allergen from olive (Olea europaea L.) pollen, increases its expression and is released to the culture medium during in vitro germination. Plant cell physiol. 2004;45(9):1149-57.
15. Barderas, R, Purohit, A, Rodriguez, R, Pauli, G and Villalba, M. Isolation of the main allergen Fra e 1 from ash (Fraxinus excelsior) pollen: comparison of the natural and recombinant forms. Ann Allergy Asthma Immunol. 2006;96(4):557-63.
16. Batanero, E, Gonzalez De La Pena, MA, Villalba, M, Monsalve, RI, Martin-Esteban, M and Rodriguez, R. Isolation, cDNA cloning and expression of Lig v 1, the major allergen from privet pollen. Clin Exp Allergy. 1996;26(12):1401-10.
17. Gonzalez, E, Villalba, M and Rodriguez, R. Immunological and molecular characterization of the major allergens from lilac and privet pollens overproduced in Pichia pastoris. Clin Exp Allergy. 2001;31(2):313-21.
18. Castro, L, Mas, S, Barderas, R, Colas, C, Garcia-Selles, J, Barber, D, et al. Sal k 5, a member of the widespread Ole e 1-like protein family, is a new allergen of Russian thistle (Salsola kali) pollen. Int Arch Allergy Immunol. 2014;163(2):142-53.
19. Barderas, R, Villalba, M, Lombardero, M and Rodriguez, R. Identification and characterization of Che a 1 allergen from Chenopodium album pollen. Int Arch Allergy Immunol. 2002;127(1):47-54.
20. Assarehzadegan, MA, Sankian, M, Jabbari, F, Noorbakhsh, R and Varasteh, A. Allergy to Salsola Kali in a Salsola incanescens-rich area: role of extensive cross allergenicity. Allergol Int. 2009;58(2):261-6.
21. Bradford, MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
22. Ali-Sadeghi, H, Khodadadi, A, Amini, A, Assarehzadegan, M-A, Sepahi, N and Zarinhadideh, F. Pro j 2 is mesquite profilin: molecular characteristics and specific IgE binding activity. Asian Pac J Allergy Immunol. 2015;33:90-8.
23. Chomczynski, P and Sacchi, N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162(1):156-9.
24. Calabozo, Bn, Dأaz-Perales, A, Salcedo, G, Barber, D and Polo, F. Cloning and expression of biologically active Plantago lanceolata pollen allergen Pla l 1 in the yeast Pichia pastoris. Biochem J. 2003;372:889-96.
25. Asturias, JA, Arilla, MC, Gomez-Bayon, N, Martinez, J, Martinez, A and Palacios, R. Cloning and expression of the panallergen profilin and the major allergen (Ole e 1) from olive tree pollen. J Allergy Clin Immunol. 1997;100(3):365-72.
26. Varasteh, A-R, Sankian, M, Midoro-Horiuti, T, Moghadam, M, Shakeri, MT, Brooks, EG, et al. Molecular cloning and expression of Cro s 1: an occupational allergen from saffron pollen (Crocus sativus). Rep Biochem Mol Biol. 2012;1(1):1-8.
27. Assarehzadegan, MA, Sankian, M, Jabbari, F, Tehrani, M and Varasteh, A. Expression of the recombinant major allergen of Salsola kali pollen (Sal k 1) and comparison with its low-immunoglobulin E-binding mutant. Allergol Int. 2010;59(2):213-22.
28. Zarinhadideh, F, Amini, A, Assarehzadegan, MA, Borsi, SH, Sepahi, N and Ali-Sadeghi, H. Immunochemical and molecular characterization of allergenic profilin (Koc s 2) from Kochia scoparia pollen. J Korean Soc Appl Biol Chem. 2015;58(3):443-51.
29. Assarehzadegan, M-A, Sankian, M, Jabbari, F, Noorbakhsh, R and Varasteh, A. Allergy to Salsola Kali in a Salsola incanescens-rich area: role of extensive cross allergenicity. Allergol Int. 2009;58(2):261-6.
30. Vahedi, F, Sankian, M, Moghadam, M, Mohaddesfar, M, Ghobadi, S and Varasteh, AR. Cloning and expression of Che a 1, the major allergen of Chenopodium album in Escherichia coli. Appl Biochem Biotech. 2011;163(7):895-905.
31. Varasteh, AR, Moghadam, M, Vahedi, F, Kermani, T and Sankian, M. Cloning and expression of the allergen Cro s 2 profilin from saffron (Crocus sativus). Allergol Int. 2009;58(3):429-35.
32. Fereidouni, M, Hossini, RF, Azad, FJ, Assarehzadegan, M-A and Varasteh, A. Skin prick test reactivity to common aeroallergens among allergic rhinitis patients in Iran. Allergol Immunopathol (Madr). 2009;37(2):73-9.
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| Issue | Vol 15, No 2 (2016) | |
| Section | Original Article(s) | |
| Keywords | ||
| Allergen Cloning Expression Prosopis | ||
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