Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (20): 3679-3682.doi: 10.3969/j.issn.1673-8225.2011.20.017

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Expression and identification of chimeric protein truncated from Ascaris allergen ABA-1

Yu Chao-sheng1, Wen Zhong1, Zou Ze-hong2, Chen Hui-fang2, Tao Ai-lin2   

  1. 1Department of Otolaryngology-Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou  510282, Guangdong Province, China
    2Academic Key Laboratory of Allergy and Clinical Immunity, State Key Laboratory of Respiratory Diseases, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou  510260, Guangdong Province, China
  • Received:2011-02-26 Revised:2011-04-09 Online:2011-05-14 Published:2011-05-14
  • Contact: Tao Ai-lin, Doctor, Professor, Master’s supervisor, Academic Key Laboratory of Allergy and Clinical Immunity, State Key Laboratory of Respiratory Diseases, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, Guangdong Province, China AerobiologiaTao@ 163. com
  • About author:Yu Chao-sheng★, Studying for master’s degree, Department of Otolaryngology- Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China ycs_9982@163.com Wen Zhong, Doctor, Professor, Department of Otolaryngology-Head and Neck Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China wenzhong60@163. com Yu Chao-sheng and Wen Zhong contributed equally to this paper.
  • Supported by:

    Great Project from the Major Program of National Science and Technology of China, No. 2008ZX08011-005*; Key Project from the Major Program of National Science and Technology of China, No. 2009ZX08011-004B*; the National Natural Science Foundation of China, No. 30771240*; Program for Educational & Scientific Innovation Team of Guangzhou City, No. B94118* 

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Abstract:

BACKGROUND: Serious allergic reaction could be caused by Ascaris anaphylactogen ABA-1 protein (Ascaris Body fluid allergen 1).
OBJECTIVE: To express the chimeric protein truncated from Ascais major allergen ABA-1 in Escherichia coli.
METHODS: The chimeric gene, obtained from GeneBank and Protein Database and named as BAA hereafter, was constructed by tailoring the coding fragments of Ascaris main allergen ABA-1, i.e., ABA-1B1, ABA-1A2 and ABA-1A1. The fusion gene BAA was synthesized and cloned into expression vector PET-44a, and transformed into Escherichia coli JM109 with the method of KCM .When the plasmid was confirmed by double restriction enzymatic digestive test with NdeⅠand PstⅠand DNA sequencing, it was transformed into Escherichia coli RosettaBlueTM step by step. The produced chimeric protein BAA was purified by nickel ion affinity chromatography, followed by identification by Western Blot and protein sequencing.
RESULTS AND CONCLUSION: Chimeric protein was inducibly expressed in Escherichia coli, with the production up to 40% of the total protein. Protein purity could reach about 90% after purification. Western Blot showed a specific band at 45 000, and amino acid sequencing identified that the 15 N-terminal amino acids were identical to the target protein. The chimeric protein has been efficiently expressed in Escherichia coli.

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