in silico design of truncated omp31 protein of brucella melitensis: its cloning and high level expression in escherichia coli
نویسندگان
چکیده
introduction: omp31 is animmunodominant and protective antigen conserved in brucella species and a good candidate for vaccine design. material & methods: the present study aimed at in silico design of the truncated omp31 (tomp31) using bioinformatic tools and to express the selected form in escherichia coli (e. coli) results and conclusion: various bioinformatically calculated scores for the model showed that the structure conformation of the truncated omp31 is in the range of the native protein with the c-score, z-score, tm-score and ramachandran z-score of the truncated form model being -0.53, -0.72 and -0.98 respectively. amplification of tomp31 produced a single fragment of approximately 345 bp which was cloned in the pet28a expression vector. the integrity of the constructed vector (pet28- tomp31) was confirmed by pcr and restriction digestion analysis and the rtomp31 was successfully expressed in e. coli bl21. sds-page analysis of the lysate from the induced e. coli carrying the tomp31 construct and the purified protein showed the expected molecular mass of approximately 12 kda. the yield of the purified protein was estimated at approximately 250 μg/ml. anti-his antibody reacted with the purified protein in western blot confirming its expression in the prokaryotic system. future studies exploring the immunogenicity and cross-protection of the protein against brucella spp. are underway. vac res , 2014, 1 (1): 16-21
منابع مشابه
In silico Design of Truncated Omp31 Protein of Brucella melitensis: Its Cloning and High Level Expression in Escherichia coli
Introduction: Omp31 is animmunodominant and protective antigen conserved in Brucella species and a good candidate for vaccine design. Material & methods: The present study aimed at in silico design of the truncated Omp31 (TOmp31) using bioinformatic tools and to express the selected form in Escherichia coli (E. coli) Results and conclusion: Various bioinformatically calculated scores for the ...
متن کاملIn silico cloning and bioinformatics study of Brucella melitensis Omp31 antigen in different mammalian expression vectors
Brucella melitensis, as a pathogenic gram-negative intracellular bacterium, causes brucellosis in animals and humans. According to literature, the B. melitensis outer membrane protein 31 (Omp31) is considered as an important vaccine candidate against brucellosis. The aim of the current study was to compare three different expression constructs containing B. melitensis Omp31 antigen using bioinf...
متن کاملIsolation, cloning and expression of the Brucella melitensis Omp31 gene
Brucellosis is a zoonotic disease transmitted to humans either from animals or from their products.Although brucellosis can be found worldwide, the Mediterranean Basin, South and Central America, EasternEurope, Asia, Africa, the Caribbean, and the Middle East have currently been listed as high-risk regions. Thegenus Brucella is classified in at least nine species. Brucella melitensis is the glo...
متن کاملin silico design, cloning and high level expression of l7/l12-tomp31 fusion protein of brucella antigens
globally, brucella melitensis and b. abortus are the most common cause of human brucellosis. the outer membrane protein 31 (omp31) and l7/l12 are immunodominant and protective antigens conserved in human brucella pathogens which are considered as potential vaccine candidates. we aimed to design the fusion protein from brucella l7/l12 and truncated omp31proteins, in silico , clone the fusion in ...
متن کاملisolation, cloning and expression of the brucella melitensis omp31 gene
brucellosis is a zoonotic disease transmitted to humans either from animals or from their products.although brucellosis can be found worldwide, the mediterranean basin, south and central america, easterneurope, asia, africa, the caribbean, and the middle east have currently been listed as high-risk regions. thegenus brucella is classified in at least nine species. brucella melitensis is the glo...
متن کاملCloning and expression of Brucella outer membrane protein 36kDa (OMP2b) in E. coli
Background & Objective: Brucellosis is an important zoonotic disease of economic significance. Brucella species are gram-negative, facultative intracellular bacteria, and are capable of replicating in the phagosomes of macrophages. They cause infection in several animal species and humans. Prevention of new diseases and diagnosis of cases infected with the organism are both essential for eradic...
متن کاملمنابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
vaccine researchجلد ۱، شماره ۱، صفحات ۱۶-۲۰
کلمات کلیدی
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023