Presentation is loading. Please wait.

Presentation is loading. Please wait.

Isolation and characterization of a clone encoding a major allergen (Bla g Bd90K) involved in IgE-mediated cockroach hypersensitivity  Ricki Helm, PhDa,

Published byLouisa Fowler Modified over 5 years ago

Similar presentations

Presentation on theme: "Isolation and characterization of a clone encoding a major allergen (Bla g Bd90K) involved in IgE-mediated cockroach hypersensitivity  Ricki Helm, PhDa,"— Presentation transcript:

1 Isolation and characterization of a clone encoding a major allergen (Bla g Bd90K) involved in IgE-mediated cockroach hypersensitivity  Ricki Helm, PhDa, Gael Cockrell, BSa, J.Steven Stanley, PhDb, Richard J. Brenner, PhDc, Wesley Burks, MDa, Gary A. Bannon, PhDb  Journal of Allergy and Clinical Immunology  Volume 98, Issue 1, Pages (July 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 SDS-PAGE (10%) and Western blot analysis of whole body German cockroach extract. Panel A, Coomassie blue–stained gel of whole body German cockroach extract. Panel B, Autoradiograph of IgE immunoblot of whole body German cockroach extract with a serum pool from eight cockroach-sensitive individuals, followed by radiolabeled anti-IgE. Lane 1, Molecular weight markers; lane 2, whole body extract; TD, tracking dye front. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 2 Selected cockroach cDNA clones encode a variety of different molecular weight IgE-binding proteins. Representative autoradiograph of a 10% SDS-PAGE/immunoblot of five cDNA clones expressing IgE-binding proteins detected by radiolabeled anti-IgE. Lanes 1-5 represent separately isolated cockroach cDNA clones. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 3 The IgE-selected cockroach clones have highly similar nucleotide sequences. Dideoxythymidine DNA sequencing reactions were performed on the 3’ end of each clone with the M13 forward primer. Each lane (A-J) represents a different IgE-selected cDNA clone. Arrowheads are reference points after the poly A tail, which indicate where each of the clones have identical patterns. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 4 Northern blot analysis of Bla g 90 kd. Cockroach poly A+ RNA was isolated from B. germanica species, and 3 μg was electrophoresed on denaturing formaldehyde agarose gels. A purified, α-32P-deoxycytidine triphosphate–labeled 4 kb insert from clone Bla g 90 kd was used as hybridization probe of a Northern blot of this gel. Lane 1, RNA ladder expressed in kilobases; lane 2, total RNA; lane 3, poly A+ RNA. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 5 Nucleotide sequence of a cockroach allergen. The nucleotide sequence of Bla g 90 kd is shown. Boxed and bold nucleotides represent one tandem repeat present in this gene. A total of seven, highly homologous (~90%) tandem repeats are contained within this cDNA clone. Numbers on the left indicate the position of the nucleotide sequence relative to the 5’ end (nucleotide position 1) and 3’ end (nucleotide position 4125) of the gene. The entire sequence has been deposited in the GenBank database, accession no. L47595. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

7 FIG. 6 Autoradiographic photographs of 10% SDS-PAGE/immunoblot analysis of German cockroach proteins incubated with serum from cockroach-sensitive, atopic, and nonatopic individuals, followed by radiolabeled anti-IgE. Panel A, Representative IgE immunoblot of a crude whole body German cockroach extract. Panel B, Representative IgE immunoblot of Bla g 90 kd expressed protein. Panels A and B: Lane 1, Pooled serum from eight cockroach-sensitive individuals; lanes 2-11, individual serum samples from known cockroach-sensitive individuals. Exposure time = 24 hours. Panel C, Representative IgE immunoblot of Bla g 90 kd expressed protein with control serum. Lane 1, Whole body extract incubated with pooled cockroach-sensitive serum; lanes 2-6, Bla g 90 kd expressed in E. coli cells; lane 2, serum from a known cockroach-sensitive individual; lane 3, nonatopic serum; lane 4, pooled serum from eight mite-sensitive individuals; lane 5, pooled serum from Alternaria-sensitive individuals; lane 6, atopic individual with high IgE titer, not cockroach-sensitive. Exposure time = 5 days. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

8 FIG. 6 Autoradiographic photographs of 10% SDS-PAGE/immunoblot analysis of German cockroach proteins incubated with serum from cockroach-sensitive, atopic, and nonatopic individuals, followed by radiolabeled anti-IgE. Panel A, Representative IgE immunoblot of a crude whole body German cockroach extract. Panel B, Representative IgE immunoblot of Bla g 90 kd expressed protein. Panels A and B: Lane 1, Pooled serum from eight cockroach-sensitive individuals; lanes 2-11, individual serum samples from known cockroach-sensitive individuals. Exposure time = 24 hours. Panel C, Representative IgE immunoblot of Bla g 90 kd expressed protein with control serum. Lane 1, Whole body extract incubated with pooled cockroach-sensitive serum; lanes 2-6, Bla g 90 kd expressed in E. coli cells; lane 2, serum from a known cockroach-sensitive individual; lane 3, nonatopic serum; lane 4, pooled serum from eight mite-sensitive individuals; lane 5, pooled serum from Alternaria-sensitive individuals; lane 6, atopic individual with high IgE titer, not cockroach-sensitive. Exposure time = 5 days. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

9 FIG. 6 Autoradiographic photographs of 10% SDS-PAGE/immunoblot analysis of German cockroach proteins incubated with serum from cockroach-sensitive, atopic, and nonatopic individuals, followed by radiolabeled anti-IgE. Panel A, Representative IgE immunoblot of a crude whole body German cockroach extract. Panel B, Representative IgE immunoblot of Bla g 90 kd expressed protein. Panels A and B: Lane 1, Pooled serum from eight cockroach-sensitive individuals; lanes 2-11, individual serum samples from known cockroach-sensitive individuals. Exposure time = 24 hours. Panel C, Representative IgE immunoblot of Bla g 90 kd expressed protein with control serum. Lane 1, Whole body extract incubated with pooled cockroach-sensitive serum; lanes 2-6, Bla g 90 kd expressed in E. coli cells; lane 2, serum from a known cockroach-sensitive individual; lane 3, nonatopic serum; lane 4, pooled serum from eight mite-sensitive individuals; lane 5, pooled serum from Alternaria-sensitive individuals; lane 6, atopic individual with high IgE titer, not cockroach-sensitive. Exposure time = 5 days. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

Download ppt "Isolation and characterization of a clone encoding a major allergen (Bla g Bd90K) involved in IgE-mediated cockroach hypersensitivity  Ricki Helm, PhDa,"

Similar presentations

Food allergy to honey: Pollen or bee products?

Food allergy to honey: Pollen or bee products?
In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex  David I. Bernstein, MDa, Raymond.

In vivo sensitization to purified Hevea brasiliensis proteins in health care workers sensitized to natural rubber latex  David I. Bernstein, MDa, Raymond.
Identification of sesame seed allergens by 2-dimensional proteomics and Edman sequencing: Seed storage proteins as common food allergens  Kirsten Beyer,

Identification of sesame seed allergens by 2-dimensional proteomics and Edman sequencing: Seed storage proteins as common food allergens  Kirsten Beyer,
Isolation and characterization of a novel 98-kd Dermatophagoides farinae mite allergen  Lai-Chen Tsai, BSca b, Pei-Ling Chao, MSa, Horng-Der Shen, PhDa.

Isolation and characterization of a novel 98-kd Dermatophagoides farinae mite allergen  Lai-Chen Tsai, BSca b, Pei-Ling Chao, MSa, Horng-Der Shen, PhDa.
IgE-mediated reaction to a banana-flavored drug additive

IgE-mediated reaction to a banana-flavored drug additive
Allergy to latex, avocado pear, and banana: Evidence for a 30 kd antigen in immunoblotting  François Lavaud, MD, Alain Prevost, MD, Claude Cossart, MD,

Allergy to latex, avocado pear, and banana: Evidence for a 30 kd antigen in immunoblotting  François Lavaud, MD, Alain Prevost, MD, Claude Cossart, MD,
Identification and molecular characterization of Charybdis feriatus tropomyosin, the major crab allergen  Patrick S.C. Leung, PhDa, Yen-chen Chen, MSca,

Identification and molecular characterization of Charybdis feriatus tropomyosin, the major crab allergen  Patrick S.C. Leung, PhDa, Yen-chen Chen, MSca,
The 14.6 kd rubber elongation factor (Hev b 1) and 24 kd (Hev b 3) rubber particle proteins are recognized by IgE from patients with spina bifida and.

The 14.6 kd rubber elongation factor (Hev b 1) and 24 kd (Hev b 3) rubber particle proteins are recognized by IgE from patients with spina bifida and.
Lupine allergy: Not simply cross-reactivity with peanut or soy

Lupine allergy: Not simply cross-reactivity with peanut or soy
Wheat α-amylase inhibitor: A second route of allergic sensitization

Wheat α-amylase inhibitor: A second route of allergic sensitization
Christopher L. Kepley, PhD, Bridget S. Wilson, PhD, Janet M

Christopher L. Kepley, PhD, Bridget S. Wilson, PhD, Janet M
Angel Vallverdú, BSc, Juan A. Asturias, PhD, M

Angel Vallverdú, BSc, Juan A. Asturias, PhD, M
Terumi Midoro-Horiuti, MD, PhDa, Randall M

Terumi Midoro-Horiuti, MD, PhDa, Randall M
Food allergy to honey: Pollen or bee products?

Food allergy to honey: Pollen or bee products?
Gloria García-Casado, PhD, Jesús F

Gloria García-Casado, PhD, Jesús F
Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant

Skin-Specific Expression of ank-393, a Novel Ankyrin-3 Splice Variant
Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,

Signal transduction pathways triggered by the FcϵRIIb receptor (CD23) in human monocytes lead to nuclear factor-κB activation  Rosa M. Ten, MD, PhDa,
Digestibility of allergens extracted from natural rubber latex and vegetable foods  Takeshi Yagami, PhDa, Yuji Haishima, PhDa, Akitada Nakamura, PhDa,

Digestibility of allergens extracted from natural rubber latex and vegetable foods  Takeshi Yagami, PhDa, Yuji Haishima, PhDa, Akitada Nakamura, PhDa,
Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen  Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo.

Molecular cloning, expression, and characterization of a major 38-kd cochineal allergen  Yoko Ohgiya, MS, Fumihiro Arakawa, MS, Hiroshi Akiyama, PhD, Yasuo.
Sabine Fischer, MSc,a, Monika Grote, PhD,b, B. Fahlbusch, PhD,c, W. D

Sabine Fischer, MSc,a, Monika Grote, PhD,b, B. Fahlbusch, PhD,c, W. D

Similar presentations

Ads by Google