مقالههای A Jahanian
توجه: محتویات این صفحه به صورت خودکار پردازش شده و مقالههای نویسندگانی با تشابه اسمی، همگی در بخش یکسان نمایش داده میشوند.
اطلاعات انتشار: Research in Pharmaceutical Sciences، دهم،شماره۱(پياپي ۲۸)، ۲۰۱۵، سال ۰
تعداد صفحات: ۶
Yersinia pestis which is the causative agent of pneumonic plague and distributed in all continents has led to many deaths during the history. Because of its high mortality rate, it must be diagnosed and treated at the earliest time post infection and therefore, rapid diagnostic tests are required. In the present study, we cloned the coding sequence of F1 capsular antigen of the bacteria in the pBAD\gIII plasmid for later expression and purification of the protein to produce poly and monoclonal antibodies against this antigen, and subsequently to develop rapid and efficient diagnostics tools for Y. pestis infections.
۲In silico design, cloning and high level expression of L7\L12–TOmp31 fusion protein of Brucella antigens
نویسنده(ها): M Golshani، s Rafati، A Jahanian، Najafabadi، M Nejati، Moheimani، SD Siadat، F Shahcheraghi، S Bouzari
اطلاعات انتشار: Research in Pharmaceutical Sciences، دهم،شماره۵(پياپي ۳۲)، ۲۰۱۵، سال ۰
تعداد صفحات: ۱۰
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 pET28a vector, and express it in Escherichia coli host. Two possible fusion forms, L7\L12–TOmp31 and TOmp31–L7\L12 were subjected to in silico modeling and analysis. Analysis and validation of the fusion proteins with three dimensional (3D) models showed that both models are in the range of native proteins. However, L7\L12–Tomp31 structure was more valid than the TOmp31 L7\L12 model and subjected to in vitro production. The major histocompatibility complex (MHC II) epitope mapping using IEDB database indicated that the model contained good MHC II binders. The L7\L12–TOmp31 coding sequence was cloned in pET28a vector. The integrity of the construct was confirmed by polymerase chain reaction, restriction enzyme mapping, and sequencing. The fusion was successfully expressed in E. coli BL21 (DE3) by induction with isopropyl β–D–thiogalactopyranoside. The rL7\L12–TOmp31 was purified with Ni–NTA column. The yield of the purified rL7\L12–TOmp31 was estimated by Bradford method and found to be 40 mg\L of the culture. Western blotting with anti–His antibody revealed a specific reactivity with purified rL7\L12–TOmp31 produced in E. coli and showed the functional expression in the prokaryotic system. In this study, a new protein vaccine candidate against brucellosis was constructed with the help of bioinformatics tools and the construct was expressed in the bacterial host. Studies evaluating the immunogenicity and cross–protection of this fusion protein against B. melitensis and B. abortus are underway.
نمایش نتایج ۱ تا ۲ از میان ۲ نتیجه