OPTIMIZING PRODUCTION OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR IN NON-PATHOGENIC LEISHMANIA BY TWO GENETIC CONSTRUCTS

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HEMAYATKAR MAHDI; DAVOUDI NOUSHIN; DAVAMI FATEMEH; MAJIDZADEH A. KEIVAN; BARKHORDARI FARZANEH; MAHBOUDI FEREIDOUN

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Journal: Tehran University Medical Journal Year:2011 | Volume:68 | Issue:11 Page(s): 629-636

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Title

OPTIMIZING PRODUCTION OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR IN NON-PATHOGENIC LEISHMANIA BY TWO GENETIC CONSTRUCTS

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Keywords

EXPRESSIONQ3

LEISHMANIA TARENTOLAEQ3

TISSUE PLASMINOGEN ACTIVATORQ3

T-PAQ3

Abstract

Background: Recombinant TISSUE PLASMINOGEN ACTIVATOR (rt-PA) is one of the most important thrombolytic agents used in patients with vascular occlusions such as acute ischemic stroke or myocardial infarction. A variety of recombinant protein EXPRESSION systems have been developed for heterologous gene EXPRESSION in prokaryotic and eukaryotic hosts. In recent years, LEISHMANIA TARENTOLAE (L. tarentolae), a nonpathogenic trypanosomatid protozoa, has come under consideration because of its safety and immunogenicity as a vaccine vector and special attributes in the EXPRESSION of complex proteins. This study was done to improve rt-PA EXPRESSION in this protozoon and create the opportunity for the replacement of rt-PA gene with other genes for the production of other complex proteins.Methods: Two EXPRESSION cassettes were used for the integration of two copies of T-PA cDNA, one copy in each cassette, into the parasite genome by electroporation. The transformed clones were selected by antibiotic resistancy. The EXPRESSION of active secreted rt-PA was confirmed by Western blot analysis and Chromolize assay.Results: Appearance of a 64 kD band in nitrocellulose membrane in the Western blot analysis confirmed the presence of full-length rt-PA in the culture media. Chromolize assay showed the EXPRESSION levels of active recombinant T-PA in single and double transfected L. tarentolae clones- 375 IU/ml and 480 IU/ml of the culture media, respectively.Conclusion: The produced rt-PA in the culture media containing the transfected cellswas at least seven times higher than what has been reported in previous studies on L. tarentolae or on some other eukaryotic systems.

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APA: Copy

HEMAYATKAR, MAHDI, DAVOUDI, NOUSHIN, DAVAMI, FATEMEH, MAJIDZADEH A., KEIVAN, BARKHORDARI, FARZANEH, & MAHBOUDI, FEREIDOUN. (2011). OPTIMIZING PRODUCTION OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR IN NON-PATHOGENIC LEISHMANIA BY TWO GENETIC CONSTRUCTS. TEHRAN UNIVERSITY MEDICAL JOURNAL (TUMJ), 68(11), 629-636. SID. https://sid.ir/paper/38618/en

Vancouver: Copy

HEMAYATKAR MAHDI, DAVOUDI NOUSHIN, DAVAMI FATEMEH, MAJIDZADEH A. KEIVAN, BARKHORDARI FARZANEH, MAHBOUDI FEREIDOUN. OPTIMIZING PRODUCTION OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR IN NON-PATHOGENIC LEISHMANIA BY TWO GENETIC CONSTRUCTS. TEHRAN UNIVERSITY MEDICAL JOURNAL (TUMJ)[Internet]. 2011;68(11):629-636. Available from: https://sid.ir/paper/38618/en

IEEE: Copy

MAHDI HEMAYATKAR, NOUSHIN DAVOUDI, FATEMEH DAVAMI, KEIVAN MAJIDZADEH A., FARZANEH BARKHORDARI, and FEREIDOUN MAHBOUDI, “OPTIMIZING PRODUCTION OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR IN NON-PATHOGENIC LEISHMANIA BY TWO GENETIC CONSTRUCTS,” TEHRAN UNIVERSITY MEDICAL JOURNAL (TUMJ), vol. 68, no. 11, pp. 629–636, 2011, [Online]. Available: https://sid.ir/paper/38618/en

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