Transformation of Nonselectable Reporter Genes in Marine Diatoms View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1999-05

AUTHORS

Angela Falciatore, Raffaella Casotti, Catherine Leblanc, Chiara Abrescia, Chris Bowler

ABSTRACT

: We report the genetic transformation of two marine diatoms by microparticle bombardment. The pennate diatom Phaeodactylum tricornutum was transformed with the bacterial gene Sh ble from Streptoalloteichus hindustanus, which confers resistance to the antibiotics phleomycin and zeocin. Transformants contained between 1 and 10 copies of the exogenous DNA integrated into the genome by illegitimate recombination at apparently random locations. Transformation efficiencies were around 10(-6), and individual cell lines could be maintained at -80 degrees C following cryopreservation. Also, P. tricornutum could be transformed simultaneously with two different plasmids, one containing the Sh ble gene and another containing the firefly luciferase gene (LUC) under control of a promoter derived from a fucoxanthin, chlorophyll a/c-binding protein gene (FCP). In these cotransformants, LUC activity was light inducible. The transient transformation of the centric diatom Thalassiosira weissflogii with the bacterial beta-glucuronidase (GUS) gene has also been achieved using similar transformation technology. The availability of gene transfer protocols for marine diatoms, together with a range of functional reporter genes and regulated expression systems, will permit molecular dissection of their biology and allow an assessment of the biotechnological potential of these organisms. More... »

PAGES

239-251

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/pl00011773

DOI

http://dx.doi.org/10.1007/pl00011773

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1011070273

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/10383998


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