IN SILICO DESIGN, CLONING AND HIGH LEVEL EXPRESSION OF L7/L12-TOMP31 FUSION PROTEIN OF BRUCELLA ANTIGENS

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

GOLSHANI MARYAM; RAFATI SIMA; JAHANIAN NAJAFABADI ALI; NEJATI MOHEIMANI MEHDI; SIADAT SEYED DAVAR; SHAHCHERAGHI FERESHTEH; BOUZARI SAEID

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: RESEARCH IN PHARMACEUTICAL SCIENCES (RPS) Year:2015 | Volume:10 | Issue:5 Page(s): 436-445

Download Full-Text

View:

276

Download:

120

Cites:

Information Journal Paper

Title

IN SILICO DESIGN, CLONING AND HIGH LEVEL EXPRESSION OF L7/L12-TOMP31 FUSION PROTEIN OF BRUCELLA ANTIGENS

Author(s)

Keywords

BRUCELLA

OMP31

L7/L12

FUSION PROTEIN

CLONING

IN SILICO DESIGN

Abstract

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 b-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.

Cites

No record.

References

No record.

Cite

APA: Copy

GOLSHANI, MARYAM, RAFATI, SIMA, JAHANIAN NAJAFABADI, ALI, NEJATI MOHEIMANI, MEHDI, SIADAT, SEYED DAVAR, SHAHCHERAGHI, FERESHTEH, & BOUZARI, SAEID. (2015). IN SILICO DESIGN, CLONING AND HIGH LEVEL EXPRESSION OF L7/L12-TOMP31 FUSION PROTEIN OF BRUCELLA ANTIGENS. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), 10(5), 436-445. SID. https://sid.ir/paper/318407/en

Vancouver: Copy

GOLSHANI MARYAM, RAFATI SIMA, JAHANIAN NAJAFABADI ALI, NEJATI MOHEIMANI MEHDI, SIADAT SEYED DAVAR, SHAHCHERAGHI FERESHTEH, BOUZARI SAEID. IN SILICO DESIGN, CLONING AND HIGH LEVEL EXPRESSION OF L7/L12-TOMP31 FUSION PROTEIN OF BRUCELLA ANTIGENS. RESEARCH IN PHARMACEUTICAL SCIENCES (RPS)[Internet]. 2015;10(5):436-445. Available from: https://sid.ir/paper/318407/en

IEEE: Copy

MARYAM GOLSHANI, SIMA RAFATI, ALI JAHANIAN NAJAFABADI, MEHDI NEJATI MOHEIMANI, SEYED DAVAR SIADAT, FERESHTEH SHAHCHERAGHI, and SAEID BOUZARI, “IN SILICO DESIGN, CLONING AND HIGH LEVEL EXPRESSION OF L7/L12-TOMP31 FUSION PROTEIN OF BRUCELLA ANTIGENS,” RESEARCH IN PHARMACEUTICAL SCIENCES (RPS), vol. 10, no. 5, pp. 436–445, 2015, [Online]. Available: https://sid.ir/paper/318407/en

Related Journal Papers

No record.

Related Seminar Papers

No record.

Related Plans

No record.

Recommended Workshops