subcloning and expression of recombinant echinococcus granulosus antigen b, in pqe-30 expression vector

نویسندگان

n taghipour

m bandepour

r pazoki

a haghighi

چکیده

background: echinococcosis or hydatid disease is a zoonotic infection caused by larval (metacestode) stages of cestodes belonging to the genus echinococcus, family taeniidae . we aimed to subclone antigen b gene in pqe-30 plasmid, its expression, and purification. methods: we subcloned hi gene into pqe-30 expression vector. the recombinant vector was transformed into e. coli , m15 and mass cultured. the subcloned gene was expressed by iptg. subcloning of gene was confirmed by both pcr and enzyme digestion. results: production of recombinant protein was confirmed by sds-page. western blot analysis was carried out by both his-tag monoclonal ab and human serum to estimate the expressed protein in e. coli cells. recombinant protein was purified and its specificity was proved by western blotting. conclusion: production of this recombinant protein can increase sensitivity and specificity in serological test (elisa).

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

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

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

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

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

Construction and Eukaryotic Expression of Recombinant Large Hepatitis Delta Antigen

Background: Hepatitis delta virus (HDV) is a subviral human pathogen that exploits host RNA editing activity to produce two essential forms of the sole viral protein, hepatitis delta antigen (HDAg). Editing at the amber/W site of HDV antigenomic RNA leads to the production of the large form (L-HDAg), which is required for RNA packaging. Methods: In this study, PCR-based site-directed mutagen...

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 parasitology

جلد ۴، شماره ۴، صفحات ۱-۹

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023