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Mem. Inst. Oswaldo Cruz ; 112(2): 155-159, Feb. 2017. tab, graf
Article in English | LILACS | ID: biblio-1040566

ABSTRACT

BACKGROUND Gene reporter-fluorescent cells have emerged as alternative method for drug screening. OBJECTIVE Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. METHODS L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. FINDINGS Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. MAIN CONCLUSION Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.


Subject(s)
Animals , Leishmania braziliensis/drug effects , Amphotericin B/pharmacology , Green Fluorescent Proteins/pharmacology , Drug Evaluation, Preclinical/methods , Antiprotozoal Agents/pharmacology , Time Factors , Microscopy, Fluorescence
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