Volume 56, Issue 1 (30 1998)
Tehran Univ Med J 1998, 56(1): 1-8 |
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Doosty M, Shirvany H. Isolation and Identification of Outer Membrane Proteins of Helicobacter Pylori of Iranian Patient by SDS-PAGE. Tehran Univ Med J 1998; 56 (1) :1-8
URL: http://tumj.tums.ac.ir/article-1-1585-en.html
URL: http://tumj.tums.ac.ir/article-1-1585-en.html
Abstract: (9298 Views)
The function of Helicobacter pylori (H.pylori) is confirmed as one of the factors which motivates gastric and duodenal ulcer and gastritis. Various methods are used to diagnose the infection. Serological tests are the easiest and most harmless for the patients. Probably, H.pylori strains in Iran are different from the strains in other countries. Hence, it seems neccessary to design a specific serological test to recognize and identify different strains of bacterial antigenic proteins of Iranian patients.
Since the most manifest and specific to these bacterial antigens are the "Outer Membrane Protein" (OMP), therefore, the first necessary step is to separate and purify H.pylori OMP and then to identify antigenic proteins.
In this study, we received bacteria colony that belonged to 15 patients with gastric or duodenal ulcer, which had been growed in blood agar or brucella broth. After processing such as washing, freezing and defreezing, sonicating, centrifugation with high speed (10,000 g) and treatment with sarcosyl, the sarcosyl insoluble fraction was extracted. Sodium Dodecyl Sulfate - Poly Acrylamide Gel Electrophoresis (SDS-PAGE) was preformed. From all 15 OMP specimens, we isolated protein bands.
The first two bands with higher MW, were major bands and the two lighter bands were the minor bands. Approximate MW of these 4 proteins are equal to 67000, 61000, 30000 and 17000 dalton
Since the most manifest and specific to these bacterial antigens are the "Outer Membrane Protein" (OMP), therefore, the first necessary step is to separate and purify H.pylori OMP and then to identify antigenic proteins.
In this study, we received bacteria colony that belonged to 15 patients with gastric or duodenal ulcer, which had been growed in blood agar or brucella broth. After processing such as washing, freezing and defreezing, sonicating, centrifugation with high speed (10,000 g) and treatment with sarcosyl, the sarcosyl insoluble fraction was extracted. Sodium Dodecyl Sulfate - Poly Acrylamide Gel Electrophoresis (SDS-PAGE) was preformed. From all 15 OMP specimens, we isolated protein bands.
The first two bands with higher MW, were major bands and the two lighter bands were the minor bands. Approximate MW of these 4 proteins are equal to 67000, 61000, 30000 and 17000 dalton
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