pELMO, an optimised in-house cloning vector

Andrea E. Ramos, Claudia Marina Muñoz, Darwin A. Moreno-Pérez, Manuel A. Patarroyo

Resultado de la investigación: Contribución a RevistaArtículo

3 Citas (Scopus)

Resumen

DNA cloning is an essential tool regarding DNA recombinant technology as it allows the replication of foreign DNA fragments within a cell. pELMO was here constructed as an in-house cloning vector for rapid and low-cost PCR product propagation; it is an optimally designed vector containing the ccdB killer gene from the pDONR 221 plasmid, cloned into the pUC18 vector’s multiple cloning site (Thermo Scientific). The ccdB killer gene has a cleavage site (CCC/GGG) for the SmaI restriction enzyme which is used for vector linearisation and cloning blunt-ended products. pELMO transformation efficiency was evaluated with different sized inserts and its cloning efficiency was compared to that of the pGEM-T Easy commercial vector. The highest pELMO transformation efficiency was observed for ~500 bp DNA fragments; pELMO vector had higher cloning efficiency for all insert sizes tested. In-house and commercial vector cloned insert reads after sequencing were similar thus highlighting that sequencing primers were designed and localised appropriately. pELMO is thus proposed as a practical alternative for in-house cloning of PCR products in molecular biology laboratories.

Idioma originalEnglish (US)
Número de artículo26
PublicaciónAMB Express
Volumen7
N.º1
DOI
EstadoPublished - dic 1 2017

Huella dactilar

Genetic Vectors
Organism Cloning
Polymerase Chain Reaction
Recombinant DNA
DNA
DNA Replication
Genes
Molecular Biology
Plasmids
Technology
Costs and Cost Analysis
Enzymes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Applied Microbiology and Biotechnology

Citar esto

Ramos, Andrea E. ; Muñoz, Claudia Marina ; Moreno-Pérez, Darwin A. ; Patarroyo, Manuel A. / pELMO, an optimised in-house cloning vector. En: AMB Express. 2017 ; Vol. 7, N.º 1.
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pELMO, an optimised in-house cloning vector. / Ramos, Andrea E.; Muñoz, Claudia Marina; Moreno-Pérez, Darwin A.; Patarroyo, Manuel A.

En: AMB Express, Vol. 7, N.º 1, 26, 01.12.2017.

Resultado de la investigación: Contribución a RevistaArtículo

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