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Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen  Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo.

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Presentation on theme: "Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen  Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo."— Presentation transcript:

1 Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen 
Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo Yoshioka, PhD, Yoshiki Hayashi, PhD, Shinobu Sakai, PhD, Sumio Ito, MS, Yuko Yamakawa, MD, PhD, Satoru Ohgiya, PhD, Zenro Ikezawa, MD, PhD, Reiko Teshima, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 5, Pages e4 (May 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Immunoblots of cochineal insect extract probed with specific IgE from 3 patients' sera and control serum. Lane M, Molecular weight (MW) marker; lane C, cochineal insect extract. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Cochineal allergen 38-kd protein cDNA and amino acid sequences. Amino acids identical to those sequenced from the 4 purified proteins are shaded. The predicted signal sequence is underlined. Four putative N-linked glycosylation sites are boxed. ∗Termination codon. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Sequence alignment of cochineal allergen protein with vespid venom phospholipase allergens that showed homology by National Center for Biotechnology Information homology analysis. Residues conserved in all proteins are dark gray, and those conserved in at least 4 sequences are light gray. Insertions made to maximize alignment are indicated by a dash. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Immunoblots of cochineal insect extract and recombinant CC38K proteins probed with specific IgE from a patient's serum or control serum. Lane M, Molecular weight (MW) marker; lane C, cochineal insect extract; lane R, recombinant secreted CC38K from yeast culture medium; lane RC, Culture medium of a yeast transformant carrying the vector. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Coomassie Brilliant Blue–stained gel of cochineal insect extract
Coomassie Brilliant Blue–stained gel of cochineal insect extract. Lane M, Molecular weight (MW) marker; lane R, cochineal insect extract. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Immunoreactivity of cochineal insect extract and proteins expressed in yeast to antibody against CC38K partial peptide. Lane M, Molecular weight (MW) marker; lane C1, culture medium of a yeast transformant carrying the C1 expression plasmid; lane R, cochineal insect extract; lane C22, cell extracts of a yeast transformant carrying the CC38K C22 expression plasmid. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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