RT Journal Article
T1 Promoter Trapping in Microalgae Using the Antibiotic Paromomycin as Selective Agent
A1 Vila Spinola, Marta
A1 Díaz Santos, Encarnación
A1 Vega Naranjo, Marta de la
A1 Rodríguez, Herminia
A1 Vargas, María Ángeles
A1 León Bañares, Rosa María
AB The lack of highly active endogenous promoters to drive the expression oftransgenes is one of the main drawbacks to achieving efficient transformation of manymicroalgal species. Using the model chlorophyte Chlamydomonas reinhardtii and theparomomycin resistance APHVIII gene from Streptomyces rimosus as a marker, we havedemonstrated that random insertion of the promoterless marker gene and subsequentisolation of the most robust transformants allows for the identification of novel strongpromoter sequences in microalgae. Digestion of the genomic DNA with an enzyme thathas a unique restriction site inside the marker gene and a high number of target sites in thegenome of the microalga, followed by inverse PCR, allows for easy determination of thegenomic region, which precedes the APHVIII marker gene. In most of the transformantsanalyzed, the marker gene is inserted in intragenic regions and its expression relies on itsadequate insertion in frame with native genes. As an example, one of the new promotersidentified was used to direct the expression of the APHVIII marker gene in C. reinhardtii,showing high transformation efficiencies.
PB MDPI
SN 1660-3397
YR 2012
FD 2012-12
LK http://hdl.handle.net/10272/18231
UL http://hdl.handle.net/10272/18231
LA eng
NO Vila Spinola, M., Díaz Santos, E.,  Vega  Naranjo, M. de la, Rodríguez, H., Vargas, M. Á., & León Bañares, R. (2012). Promoter Trapping in Microalgae Using the Antibiotic Paromomycin as Selective Agent. Marine Drugs, 10(12), 2749–2765. DOI: https://doi.org/10.3390/md10122749
DS Repositorio Institucional de la Universidad de Huelva
RD 30 may 2026