molecular cloning and expression an 8-kda subunit of antigen b from g1 strain of echinococcus granulosus

نویسندگان

hakim azizi dept. of medical parasitology and mycology, school of public health, tehran university of medical sciences, iran.

bahram kazemi cellular and molecular biology research centre, shahid beheshti university of medical sciences, tehran, iran and biotechnology department, school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran.

mojgan bandehpour cellular and molecular biology research centre, shahid beheshti university of medical sciences, tehran, iran and biotechnology department, school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran.

mehdi mohebali dept. of medical parasitology and mycology, school of public health, tehran university of medical sciences, iran and center for research of endemic parasites of iran (crepi), tehran university of medical sciences, tehran, iran.

چکیده

background : echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection caused by the larval stage of the dog taeniid tapeworm echinococcus granulosus. vaccination has been considered as one of the ways to prevent of hy-datidosis in recent decades. the aim of this study was to construct a pcdna3.1 eukaryotic expression vector contain-ing the subunit 8-kda antigen b (hyd1) of e. granulosus (g1 strain) and investigate its capability to induce protein ex-pression in mammalian cell line, as a basis toward developing a dna vaccine against hydatidosis. methods : the coding sequence of hydi was amplified by pcr with the specific pcr primers from pqe/hydi, and then was sub-cloned into pcdna3.1 plasmid as expression vector. the pchyd1 plasmid was digested by restriction enzymes and amplified with the specific pcr primers to confirm cloning of this gene in pcdna3 plasmid. in last step, the sub-cloned gene was expressed in mammalian cell line (nih 3t3 cells). result : the subunit 8-kda antigen b (hyd1) was successfully sub-cloned in pcdna3.1 and hyd1 protein was ex-pressed in eukaryotic cell confirmed by sds-page and western blot. conclusion : recombinant plasmid of pcdna3.1 was successfully constructed and express of recombinant hyd1 protein was confirmed. that is promising step for forthcoming measures on providing vaccine against human and animal hydatidosis.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Molecular Cloning and Expression an 8-kDa Subunit of Antigen B from G1 strain of Echinococcus granulosus.

BACKGROUND Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection caused by the larval stage of the dog taeniid tapeworm Echinococcus granulosus. Vaccination has been considered as one of the ways to prevent of hydatidosis in recent decades. The aim of this study was to construct a pcDNA3.1 eukaryotic expression vector containing the subunit 8-kDa antigen B (Hyd1) of E. granul...

متن کامل

Gene cloning, expression and serological evaluation of the 12-kDa antigen-B subunit from Echinococcus granulosus.

A 12-kDa subunit of antigen B from Echinococcus granulosus has recently been cloned, expressed and used in diagnostic ELISA to test human sera for evidence of cystic echinococcosis. The performance of the ELISA based on the recombinant antigen (rAgB) was compared with that of similar assays based on native antigen B (nAgB) or hydatid-cyst fluid. For the preparation of the rAgB, total RNA was ex...

متن کامل

Molecular cloning, expression, and serological evaluation of an 8-kilodalton subunit of antigen B from Echinococcus multilocularis.

Full-length cDNA and genomic DNA encoding an 8-kDa subunit of antigen B from Echinococcus multilocularis (designated EmAgB8/1) were isolated from an E. multilocularis metacestode cDNA library and a protoscolex genomic DNA library, respectively. The open reading frame of the cDNA clone encodes a polypeptide comprising 85 amino acids with a 20-amino-acid NH(2)-terminal signal sequence, which was ...

متن کامل

Immunodiagnostic Value of Echinococcus Granulosus Recombinant B8/1 Subunit of Antigen B

Background: Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. Objective: The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodi...

متن کامل

Echinococcus granulosus Antigen B Structure: Subunit Composition and Oligomeric States

BACKGROUND Antigen B (AgB) is the major protein secreted by the Echinococcus granulosus metacestode and is involved in key host-parasite interactions during infection. The full comprehension of AgB functions depends on the elucidation of several structural aspects that remain unknown, such as its subunit composition and oligomeric states. METHODOLOGY/PRINCIPAL FINDINGS The subunit composition...

متن کامل

Isolation and purification of Echinococcus granulosus antigen B from hydatid cyst fluid using three different methods

Hydatid cyst, the larval stage of cestodes Echinococcus spp., is recognized as a zoonotic infection in the world. The World Health Organization (WHO) has recently classified echinococcosis in a group of neglected tropical diseases.The prevalence of Echinococcus granulosus infection is high in Iran due to the presence of various intermediate hosts in this country. Considering the rising trend of...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
iranian journal of public health

جلد ۴۴، شماره ۷، صفحات ۹۶۲-۹۶۸

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023