Achievement of constitutive fluorescent pLEXSY-egfp Leishmania braziliensis and its application as an alternative method for drug screening in vitro.
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.


RESULTS
Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin.


CONCLUSIONS
Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Matheus Silva E
MS
Bastos
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Luciana Ângelo de
LÂ
Souza
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Thiago Souza
TS
Onofre
Universidade Federal de Viçosa, Departamento de Veterinária, Viçosa, MG, Brasil.
Abelardo
A
Silva
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Márcia Rogéria de
MR
Almeida
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Gustavo Costa
GC
Bressan
Universidade Federal de Viçosa, Departamento de Bioquímica e Biologia Molecular, Viçosa, MG, Brasil.
Juliana Lopes Rangel
JL
Fietto
D000666:Amphotericin B / Q000494:pharmacology*; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000494:pharmacology*; D007892:Leishmania braziliensis / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D013997:Time Factors
D000981:Antiprotozoal Agents; D049452:Green Fluorescent Proteins; D000666:Amphotericin B