Cloning rhoptry protein 1 (ROP1) gene of Toxoplasma gondii (RH) in expression vector

Authors

  • F. Ghaffarifar
  • Z. Eslamirad
  • Z. Sharifi
Abstract:

  Toxoplasma gondii contain various immunogenic antigens. The most important Toxoplasma antigens are somatic and excreted/secreted antigens. Rhoptry proteins are known as excreted/secreted antigens. These antigens have been proposed as a vaccine candidate against toxoplasmosis. The main objective of the present work was cloning rhoptry protein1 (ROP1) Gene of Toxoplasma gondii (RH) in a cloning vector for gene analysis and further production of rhoptry proteins. Tachyzoites of the RH strain of T. gondii were harvested from the peritoneal fluid of mice that has been experimentally infected with the parasites. Genomic DNA was extracted by phenol- chloroform method. The ROP1 fragment amplified with specific primers. The purified PCR products were ligated between the EcoR1 and BamH1 sites of the pTZ57R/T cloning vector and transformed into Escherichia coli TG1 strain and screened by IPTG and X-Gal. The plasmid was purified and visualized under UV transilluminator. The amplified fragment was cloned in pTZ57R vector successfully. The correct orientation of the ROP1 fragment was identified by restriction enzyme analysis and sequencing of constructed plasmid. A fragment about 760bp was separated from PTZ57R following digestion and demonstrated on agarose gel electrophoresis.The sequence of this amplified gene showed homology up to 96% with target gene in GenBank database (Accession no. M71274). Recombinant plasmid of ROP1 gene was constructed. It is ready for future study.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

cloning rhoptry protein 1 (rop1) gene of toxoplasma gondii (rh) in expression vector

toxoplasma gondii contain various immunogenic antigens. the most important toxoplasma antigens are somatic and excreted/secreted antigens. rhoptry proteins are known as excreted/secreted antigens. these antigens have been proposed as a vaccine candidate against toxoplasmosis. the main objective of the present work was cloning rhoptry protein1 (rop1) gene of toxoplasma gondii (rh) in a cloning v...

full text

Evaluating the serological applications of Toxoplasma gondii rhoptry protein 1 (ROP1) antigen

Introduction: Toxoplasma gondii (T. gondii) is distributed worldwide and infects most species. The serious incidence and severe or fatal injury caused by T. gondii infection clearly indicates the necessity for the event of a vaccine. The current study goals were to evaluate serological applications of Toxoplasma gondiirhoptry protein 1 (ROP1) ...

full text

a preliminary study: expression of rhoptry protein 1 (rop1) toxoplasma gondii in prokaryote system

background toxoplasma gondii is an obligatory intracellular protozoan parasite, which infects human beings. since the current antigens used for diagnosis or vaccination are contaminated with non parasitic material in which the parasite is grown, it is tried to produce recombinant antigens to design vaccines against toxoplasmosis, or make diagnostic kits. choosing the type of antigen to produce ...

full text

بررسی کارایی عملکرد آنتی‌ژن‌نوترکیب ROP1 در تشخیص عفونت توکسوپلاسما گوندی

Introduction:Toxoplasma gondii is a ubiquitous obligate intracellular parasite with a relatively broad host range infecting both mammals and birds. Toxoplasma proteins are strong antigens that can begin strong immune reactions, among which Rhoptry protein 1 (ROP1) can be named discharging from rhoptry cell-organ. ROP1 is regarded as a competitor for recombinant vaccines against toxoplasmosis. T...

full text

evaluating the serological applications of toxoplasma gondii rhoptry protein 1 (rop1) antigen

introduction: toxoplasma gondii (t. gondii) is distributed worldwide and infects most species. the serious incidence and severe or fatal injury caused by t. gondii infection clearly indicates the necessity for the event of a vaccine. the current study goals were to evaluate serological applications of toxoplasma gondiirhoptry protein 1 (rop1) antigen. materials and methods: we created a polymer...

full text

cloning and expression of major surface antigen 1 gene of toxoplasma gondii rh strain using the expression vector pvax1 in chinese hamster ovary cells

background toxoplasmosis is an opportunistic protozoan infection with a high prevalence in a broad range of hosts infecting up to one-third of the world human population. toxoplasmosis leads to serious medical problems in immunocompromised individuals and fetuses and also induces abortion and mortality in domestic animals. therefore, there is a huge demand for the development of an effective va...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 63  issue 2

pages  11- 17

publication date 2008-07-01

By following a journal you will be notified via email when a new issue of this journal is published.

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023