CLONING, EXPRESSION AND EVALUATION OF PSEUDOMONAS AERUGINOSA EXOTOXIN A

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LANGARI JAHANGIR; GOLKAR MAJID; KHANAHMAD HOSSEIN; KARIMIPOOR MORTEZA; AREZUMAND ROGHAYEH; BEHZADI RAMAZAN; AHANGARI COHAN REZA

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Journal: JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S) Year:2014 | Volume:32 | Issue:294 Page(s): 1-9

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Title

CLONING, EXPRESSION AND EVALUATION OF PSEUDOMONAS AERUGINOSA EXOTOXIN A

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Keywords

PSEUDOMONAS AUROGINOSA EXOTOXIN AQ2

PE38Q2

CLONINGQ2

EXPRESSIONQ2

Abstract

Background: Nowadays, in many studies related to the treatment of various cancers, toxic compounds were targeted against cancer cells. One of the most effective compounds is Pseudomonas exotoxin A. The purpose of this study is to investigate the EXPRESSION, purification, and in vitro evaluation of a short form of the toxin in a prokaryotic EXPRESSION system.Methods: The short form of the toxin (PE38) was amplified by PCR using primers containing HindIII and NdeI restriction enzyme sites from pUC57-PE38. The PCR product was digested and subcloned in the pET-26b EXPRESSION vector. The EXPRESSION vector was separately transformed into the BL21 (DE3), BL21 (DE3) plys S and Rosetta E.coli strains. Recombinant bacteria were cultured and induced and the resulted PE38 protein purified using metal affinity column chromatography. The toxicity effect of PE38 protein was assessed on HUVEC and 293KDR eukaryotic cells.Findings: The gene was successfully cloned into the EXPRESSION vector and the accuracy of construct was confirmed by restriction map analysis and sequencing. The EXPRESSION was found more in BL21(DE3) than the other strains. The toxicity effect was observed at the same level for both HUVEC and 293KDR cells.Conclusion: The lethal dose of truncated toxin is more than the previous study (1000-fold), where the targeted vascular endothelial growth factor (VEGF121) was fused to the truncated toxin.

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APA: Copy

LANGARI, JAHANGIR, GOLKAR, MAJID, KHANAHMAD, HOSSEIN, KARIMIPOOR, MORTEZA, AREZUMAND, ROGHAYEH, BEHZADI, RAMAZAN, & AHANGARI COHAN, REZA. (2014). CLONING, EXPRESSION AND EVALUATION OF PSEUDOMONAS AERUGINOSA EXOTOXIN A. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), 32(294), 1-9. SID. https://sid.ir/paper/50642/en

Vancouver: Copy

LANGARI JAHANGIR, GOLKAR MAJID, KHANAHMAD HOSSEIN, KARIMIPOOR MORTEZA, AREZUMAND ROGHAYEH, BEHZADI RAMAZAN, AHANGARI COHAN REZA. CLONING, EXPRESSION AND EVALUATION OF PSEUDOMONAS AERUGINOSA EXOTOXIN A. JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S)[Internet]. 2014;32(294):1-9. Available from: https://sid.ir/paper/50642/en

IEEE: Copy

JAHANGIR LANGARI, MAJID GOLKAR, HOSSEIN KHANAHMAD, MORTEZA KARIMIPOOR, ROGHAYEH AREZUMAND, RAMAZAN BEHZADI, and REZA AHANGARI COHAN, “CLONING, EXPRESSION AND EVALUATION OF PSEUDOMONAS AERUGINOSA EXOTOXIN A,” JOURNAL OF ISFAHAN MEDICAL SCHOOL (I.U.M.S), vol. 32, no. 294, pp. 1–9, 2014, [Online]. Available: https://sid.ir/paper/50642/en

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