Stable nuclear transformation of Pandorina morum

Lerche K, Hallmann A (2014)
BMC Biotechnology 14(1).

Download
OA
Journal Article | Published | English
Abstract
Background Volvocine green algae like Pandorina morum represent one of the most recent inventions of multicellularity diverged from their unicellular relatives. The 8–16 celled P. morum alga and its close multicellular relatives constitute a model lineage for research into cellular differentiation, morphogenesis and epithelial folding, sexual reproduction and evolution of multicellularity. Pandorina is the largest and most complex organism in the volvocine lineage that still exhibits isogamous sexual reproduction. So far, molecular-biological investigations in P. morum were constricted due to the absence of methods for transformation of this species, which is a prerequisite for introduction of reporter genes and (modified) genes of interest. Results Stable nuclear transformation of P. morum was achieved using chimeric constructs with a selectable marker, a reporter gene, promoters and upstream and downstream flanking sequences from heterologous sources. DNA was introduced into the cells by particle bombardment with plasmid-coated gold particles. The aminoglycoside 3′-phosphotransferase VIII (aphVIII) gene of Streptomyces rimosus under control of an artificial, heterologous promoter was used as the selectable marker. The artificial promoter contained a tandem arrangement of the promoter of both the heat shock protein 70A (hsp70A) and the ribulose-1,5-bisphosphat-carboxylase/-oxygenase S3 (rbcS3) gene of Volvox carteri. Due to the expression of aphVIII, transformants gained up to 333-fold higher resistance to paromomycin in comparison to the parent wild-type strain. The heterologous luciferase (gluc) gene of Gaussia princeps, which was previously genetically engineered to match the nuclear codon usage of Chlamydomonas reinhardtii, was used as a co-transformed, unselectable reporter gene. The expression of the co-bombarded gluc gene in transformants and the induction of gluc by heat shock were demonstrated through bioluminescence assays. Conclusion Stable nuclear transformation of P. morum using the particle bombardment technique is now feasible. Functional expression of heterologous genes is achieved using heterologous flanking sequences from Volvox carteri and Chlamydomonas reinhardtii. The aphVIII gene of the actinobacterium S. rimosus can be used as a selectable marker for transformation experiments in the green alga P. morum. The gluc gene of the marine copepod G. princeps, expressed under control of heterologous promoter elements, represents a suitable reporter gene for monitoring gene expression or for other applications in P. morum.
Publishing Year
ISSN
PUB-ID

Cite this

Lerche K, Hallmann A. Stable nuclear transformation of Pandorina morum. BMC Biotechnology. 2014;14(1).
Lerche, K., & Hallmann, A. (2014). Stable nuclear transformation of Pandorina morum. BMC Biotechnology, 14(1).
Lerche, K., and Hallmann, A. (2014). Stable nuclear transformation of Pandorina morum. BMC Biotechnology 14.
Lerche, K., & Hallmann, A., 2014. Stable nuclear transformation of Pandorina morum. BMC Biotechnology, 14(1).
K. Lerche and A. Hallmann, “Stable nuclear transformation of Pandorina morum”, BMC Biotechnology, vol. 14, 2014.
Lerche, K., Hallmann, A.: Stable nuclear transformation of Pandorina morum. BMC Biotechnology. 14, (2014).
Lerche, Kai, and Hallmann, Armin. “Stable nuclear transformation of Pandorina morum”. BMC Biotechnology 14.1 (2014).
Main File(s)
Access Level
OA Open Access
Last Uploaded
2014-08-14 08:55:45

This data publication is cited in the following publications:
This publication cites the following data publications:

3 Citations in Europe PMC

Data provided by Europe PubMed Central.

Understanding nitrate assimilation and its regulation in microalgae.
Sanz-Luque E, Chamizo-Ampudia A, Llamas A, Galvan A, Fernandez E., Front Plant Sci 6(), 2015
PMID: 26579149
The inducible nitA promoter provides a powerful molecular switch for transgene expression in Volvox carteri.
von der Heyde EL, Klein B, Abram L, Hallmann A., BMC Biotechnol. 15(), 2015
PMID: 25888095

47 References

Data provided by Europe PubMed Central.

The bacterial phleomycin resistance gene ble as a dominant selectable marker in Chlamydomonas.
Stevens DR, Rochaix JD, Purton S., Mol. Gen. Genet. 251(1), 1996
PMID: 8628243
Algal transgenics and biotechnology
AUTHOR UNKNOWN, 2007
The bacterial paromomycin resistance gene, aphH, as a dominant selectable marker in Volvox carteri.
Jakobiak T, Mages W, Scharf B, Babinger P, Stark K, Schmitt R., Protist 155(4), 2004
PMID: 15648719
Bacterial resistance to aminoglycoside antibiotics.
Davies J, Wright GD., Trends Microbiol. 5(6), 1997
PMID: 9211644
Bioluminescence in the mesopelagic copepod, Gaussia princeps (T. Scott)
AUTHOR UNKNOWN, 1972
Cloning and characterization of new bioluminescent proteins
AUTHOR UNKNOWN, 1999
Enzymes involved in anaerobic respiration appear to play a role in Actinobacillus pleuropneumoniae virulence.
Jacobsen I, Hennig-Pauka I, Baltes N, Trost M, Gerlach GF., Infect. Immun. 73(1), 2005
PMID: 15618158
Stable nuclear transformation of Chlamydomonas using the Chlamydomonas gene for nitrate reductase.
Kindle KL, Schnell RA, Fernandez E, Lefebvre PA., J. Cell Biol. 109(6 Pt 1), 1989
PMID: 2592399
Stable nuclear transformation of Chlamydomonas reinhardtii with a Streptomyces rimosus gene as the selective marker.
Sizova IA, Lapina TV, Frolova ON, Alexandrova NN, Akopiants KE, Danilenko VN., Gene 181(1-2), 1996
PMID: 8973302
Stable nuclear transformation of the diatom Phaeodactylum tricornutum.
Apt KE, Kroth-Pancic PG, Grossman AR., Mol. Gen. Genet. 252(5), 1996
PMID: 8914518
Genetic transformation of the diatoms Cyclotella cryptica and Navicula saprophila
AUTHOR UNKNOWN, 1995
Reporter genes and highly regulated promoters as tools for transformation experiments in Volvox carteri.
Hallmann A, Sumper M., Proc. Natl. Acad. Sci. U.S.A. 91(24), 1994
PMID: 7972102
Gene replacement by homologous recombination in the multicellular green alga Volvox carteri.
Hallmann A, Rappel A, Sumper M., Proc. Natl. Acad. Sci. U.S.A. 94(14), 1997
PMID: 9207115

AUTHOR UNKNOWN, 1988

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 25031031
PubMed | Europe PMC

Search this title in

Google Scholar