Expression, purification and identification of Pla a1 in a codon-optimized Platanus pollen allergen

Liu,Yun; Sun,Xiuzhen; Wang,Guizuo; Tao,Ailin; Wu,Yuanyuan; Li,Manxiang; Shi,Hongyang; Xie,Mei

doi:10.3892/mmr.2015.3652

Expression, purification and identification of Pla a1 in a codon-optimized Platanus pollen allergen

Authors:
- Yun Liu
- Xiuzhen Sun
- Guizuo Wang
- Ailin Tao
- Yuanyuan Wu
- Manxiang Li
- Hongyang Shi
- Mei Xie
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Affiliations: Department of Respiratory Medicine, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shanxi 710004, P.R. China, National Key Laboratory of Respiratory Diseases, Allergy Research Center, Key Laboratory of Allergic Reactions and Clinical Immunology, The Second Affiliated Hospital of Guangzhou Medical College, Guangzhou, Guangdong 510260, P.R. China
Published online on: April 21, 2015 https://doi.org/10.3892/mmr.2015.3652
Pages: 2197-2202

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Abstract

The present study aimed to express, purify and identify the major allergen gene, Pla a1, in Platanus pollen. According to previous studies, the major gene sequences of the Pla a1 allergen were obtained and codon optimization and synthesis of the genome were performed using DNAStar software. Following binding of the target gene fragment and the pET‑44a vector, the JM109 cells were transfected to produce positive clones. The vectors were then transformed into Escherichia coli Rosetta cells to induce the expression of the target protein. The exogenous protein was purified using affinity chromatography and was identified by western blot analysis. Pla a1, the major allergen protein in Platanus pollen, was successfully isolated and this exogenous protein was purified using affinity chromatography. The present study was the first, to the best of our knowledge, to obtain expression of the allergen recombinant protein, Pla a1, fused with a Strep‑TagII via codon optimization and provided the basis for the preparation of allergens with high purity, recombinant hypoallergenic allergens and allergen nucleic acid vaccines.

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1	Kosisky SE, Marks MS and Nelson MR: Pollen aeroallergens in the Washington, DC, metropolitan area: a 10-year volumetric survey (1998–2007). Ann Allergy Asthma Immunol. 104:223–235. 2010. View Article : Google Scholar : PubMed/NCBI
2	Aira MJ, Rodriguez-Rajo FJ, Fernandez-Gonzalez M and Jato V: Airborne pollen of ornamental tree species in the NW of Spain. Environ Monit Assess. 173:765–775. 2010. View Article : Google Scholar : PubMed/NCBI
3	Ture C and Bocuk H: Analysis of airborne pollen grains in Bilecik, Turkey. Environ Monit Assess. 151:27–35. 2009. View Article : Google Scholar
4	Celenk S, Canitez Y, Bicakci A, et al: An aerobiological study on pollen grains in the atmosphere of North-West Turkey. Environ Monit Assess. 158:365–380. 2009. View Article : Google Scholar
5	Liu GH, Zhu RF, Zhang W, et al: Survey of airborne pollen in Hubei Province. Chin Med Sci J. 23:212–217. 2008. View Article : Google Scholar
6	Lu JM, Sun XZ, Liu Y, et al: Survey of airborne pollen in Xi’an City. Journal of Xi’an Jiaotong University (Medical Science Edition). 31:472–474. 2010.
7	Lauer I, Miguel-Moncin MS, Abel T, et al: Identification of a plane pollen lipid transfer protein (Pla a 3) and its immunological relation to the peach lipid-transfer protein, Pru p 3. Clin Exp Allergy. 37:261–269. 2007. View Article : Google Scholar : PubMed/NCBI
8	Alcázar P, García-Mozo H, Trigo Mdel M, et al: Platanus pollen season in Andalusia (southern Spain): trends and modeling. J Environ Monit. 13:2502–2510. 2011. View Article : Google Scholar : PubMed/NCBI
9	Mardones P, Grau M, Araya J, et al: First annual register of allergenic pollen in Talca, Chile. Allergol Immunopathol (Madr). 41:233–238. 2012. View Article : Google Scholar
10	Pérez-Badia R, Rapp A, Vaquero C and Fernández-González F: Aerobiological study in east-central Iberian Peninsula: pollen diversity and dynamics for major taxa. Ann Agric Environ Med. 18:99–111. 2011.PubMed/NCBI
11	Asturias JA, Ibarrols I, Eraso E, et al: The major Platanus acerifolia pollen allergen Pla a1 has sequence homology to invertase inhibitors. Clin Exp Allergy. 33:978–985. 2003. View Article : Google Scholar : PubMed/NCBI
12	Ibarrola I, Arilla MC, Martinez A, et al: Identification of a polygalacturonase as a major allergen (Pla a2) from Platanus acerifolia pollen. J Allergy Clin Immunol. 113:1185–1191. 2004. View Article : Google Scholar : PubMed/NCBI
13	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
14	Celenk S, Bicakci A, Tamay Z, et al: Airborne pollen in European and Asian parts of Istanbul. Environ Monit Assess. 164:391–402. 2009. View Article : Google Scholar : PubMed/NCBI
15	Subiza J, Cabrera M, Valdivieso R, et al: Seasonal asthma caused by airborne Platanus pollen. Clin Exp Allergy. 24:1123–1129. 1994. View Article : Google Scholar : PubMed/NCBI
16	Pazouki N, Sankian M, Leung PT, et al: Identification of cyclophilin as a novel allergen from Platanus orientalis pollens by mass spectrometry. J Biosci Bioeng. 107:215–217. 2009. View Article : Google Scholar : PubMed/NCBI
17	Fernández-González D, González-Parrado Z, Vega-Maray AM, et al: Platanus pollen allergen, Pla a 1: quantification in the atmosphere and influence on a sensitizing population. Clin Exp Allergy. 40:1701–1708. 2010. View Article : Google Scholar : PubMed/NCBI
18	Varela S, Subiza J, Subiza JL, et al: Platanus pollen as an important cause of pollinosis. J Allergy Clin Immunol. 100:748–754. 1997. View Article : Google Scholar
19	Baneyx F: Recombinant protein expression in Escherichia coli. Curr Opin Biotechnol. 10:411–421. 1999. View Article : Google Scholar : PubMed/NCBI
20	Zheng BQ: Escherichia coli codon preference overview. Silicon Valley. 3:242009.
21	Nikolaizik WH, Weichel M, Blaser K and Crameri R: Intracutaneous tests with recombinant allergens in cystic fibrosis patients with allergic bronchopulmonary aspergillosis and Aspergillus allergy. Am J Respir Crit Care Med. 165:916–921. 2002. View Article : Google Scholar : PubMed/NCBI