Development of a genetic tool for functional screening of anti-malarial bioactive extracts in metagenomic libraries

Malaria Journal

Methodology

Marcio M. Yamamoto¹  Silvia Restrepo²  Carmen Fernández-Becerra³  Hernando A. Del Portillo⁴  Marcela Lozano⁵  Maria M. Zambrano⁵  Patricia Del Portillo⁵  Francy J. Pérez-Llanos⁶  Alvaro Mongui⁶  Howard Junca⁷ 

[1] Departamento de Parasitologia, Universidade de São Paulo, São Paulo, Brazil;Department of Biological Sciences, Universidad de los Andes, Bogotá, Colombia;ICREA at ISGlobal, Barcelona Ctr Int Health Res (CRESIB), Hospital Clínic - Universitat de Barcelona, Barcelona, Spain;ICREA at ISGlobal, Barcelona Ctr Int Health Res (CRESIB), Hospital Clínic - Universitat de Barcelona, Barcelona, Spain;Institució Catalana de Recerca I Estudis Avançats (ICREA), Barcelona, Spain;RG Microbial Ecology: Metabolism, Genomics & Evolution - CorpoGen, Bogotá, Colombia;RG Microbial Ecology: Metabolism, Genomics & Evolution - CorpoGen, Bogotá, Colombia;Department of Biological Sciences, Universidad de los Andes, Bogotá, Colombia;RG Microbial Ecology: Metabolism, Genomics & Evolution - CorpoGen, Bogotá, Colombia;Present Address: Applied Biology Program, Faculty of Basic & Applied Sciences, Universidad Militar Nueva Granada−UMNG, Campus Cajicá, Bogotá, DC, Colombia;

关键词: Malaria;    Metagenomics;    Functional screening;    Anti-malarial drugs;    Synthetic gene design;    Dermaseptin 4;    Plasmodium falciparum;   

DOI  :  10.1186/s12936-015-0748-6

 received in 2014-10-17, accepted in 2015-05-26,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundThe chemical treatment of Plasmodium falciparum for human infections is losing efficacy each year due to the rise of resistance. One possible strategy to find novel anti-malarial drugs is to access the largest reservoir of genomic biodiversity source on earth present in metagenomes of environmental microbial communities.MethodsA bioluminescent P. falciparum parasite was used to quickly detect shifts in viability of microcultures grown in 96-well plates. A synthetic gene encoding the Dermaseptin 4 peptide was designed and cloned under tight transcriptional control in a large metagenomic insert context (30 kb) to serve as proof-of-principle for the screening platform.ResultsDecrease in parasite viability consistently correlated with bioluminescence emitted from parasite microcultures, after their exposure to bacterial extracts containing a plasmid or fosmid engineered to encode the Dermaseptin 4 anti-malarial peptide.ConclusionsHere, a new technical platform to access the anti-malarial potential in microbial environmental metagenomes has been developed.

【 授权许可】

Unknown   
© Mongui et al. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.