RT journal article
T1 New insights on expression and purification of a recombinant luciferase protein from the bioluminescence marine dinoflagellate Pyrocystis lunula
A1 Fajardo Quiñones, Carlos
A1 Fernández Acero, Francisco Javier
A1 De Donato, M.
A1 Gómez, F.
A1 Martínez Rodríguez, G.
A1 Mancera Romero, Juan Miguel
A2 Biología
A2 BiomedicinaBiotecnología y Salud Pública
K1 Pyrocystis lunula
K1 Bioluminescence
K1 Dinoflagellate
K1 Protein expression
K1 Protein purification
K1 Reporter genes
K1 Gene cloning
AB Bioluminescence is interesting, among other reasons, for the various technological applicationsthat have been derived from it. Among these applications, developing visualization techniques to record theexpression of one or more genes simultaneously in real-time are particularly useful. With this in mind, this studyaimed to generate a recombinant Pyrocystis lunula luciferase protein (Luci D2-3 partial CDS). As the mainresults, i) a fragment of 1467 bp of the luciferase (LCFb) mRNA of the dinoflagellate P. lunula, containing partof domain 2 and all of the domain 3, was cloned in the pET28a vector; ii) the constructed vector was used totransform Escherichia coli to express the recombinant protein and subsequently purify it through an affinitychromatography procedure using a His-Tag; and iii) the purified protein (~50 kDa) was further analyzed bymass spectrometry to confirm its identity. Despite being unable to perform activity tests with the luciferinsubstrate, the evidence from previous studies indicates that the recombinant protein obtained in this case isenzymatically active. Due to the limited number of currently available luciferases, synthesizing this recombinantprotein represents a useful tool, especially in designing expression assays coupled to multiple reporter genes,thus expanding the palette of proteins available for developing this type of biotechnological advances
PB Escuela de Ciencias del Mar. Pontificia Universidad Católica de Valparaiso
SN 0718-560X
YR 2023
FD 2023
LK http://hdl.handle.net/10498/31754
UL http://hdl.handle.net/10498/31754
LA eng
NO European Commission (European Union).Federación Española de Enfermedades Raras.Ministerio de Asuntos Económicos y Transformación Digital, Gobierno de España.
DS Repositorio Institucional de la Universidad de Cádiz
RD 08-nov-2026