Abstract
The fate of exogenous DNA introduced into Chlamydomonas reinhardtii by electroporation was analyzed. With single and double electrical pulses, plasmids as large as 14 kb were introduced into cells with and without intact cell walls. Within hours after introduction, exogenous plasmid DNA was associated with nuclei isolated from cells; several weeks after introduction, exogenous DNA was stably integrated into the Chlamydomonas genome. These studies establish electroporation as a method for introducing DNA, and potentially other molecules, into C. reinhardtii.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bandziulis R. J., Rosenbaum J. L. Novel control elements in the alpha-1 tubulin gene promoter from Chlamydomonas reinhardii. Mol Gen Genet. 1988 Oct;214(2):204–212. doi: 10.1007/BF00337712. [DOI] [PubMed] [Google Scholar]
- Boynton J. E., Gillham N. W., Harris E. H., Hosler J. P., Johnson A. M., Jones A. R., Randolph-Anderson B. L., Robertson D., Klein T. M., Shark K. B. Chloroplast transformation in Chlamydomonas with high velocity microprojectiles. Science. 1988 Jun 10;240(4858):1534–1538. doi: 10.1126/science.2897716. [DOI] [PubMed] [Google Scholar]
- Callis J., Fromm M., Walbot V. Expression of mRNA electroporated into plant and animal cells. Nucleic Acids Res. 1987 Jul 24;15(14):5823–5831. doi: 10.1093/nar/15.14.5823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Debuchy R., Purton S., Rochaix J. D. The argininosuccinate lyase gene of Chlamydomonas reinhardtii: an important tool for nuclear transformation and for correlating the genetic and molecular maps of the ARG7 locus. EMBO J. 1989 Oct;8(10):2803–2809. doi: 10.1002/j.1460-2075.1989.tb08426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diener D. R., Curry A. M., Johnson K. A., Williams B. D., Lefebvre P. A., Kindle K. L., Rosenbaum J. L. Rescue of a paralyzed-flagella mutant of Chlamydomonas by transformation. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5739–5743. doi: 10.1073/pnas.87.15.5739. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hasnain S. E., Manavathu E. K., Leung W. C. DNA-mediated transformation of Chlamydomonas reinhardi cells: use of aminoglycoside 3'-phosphotransferase as a selectable marker. Mol Cell Biol. 1985 Dec;5(12):3647–3650. doi: 10.1128/mcb.5.12.3647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller L. R., Schloss J. A., Silflow C. D., Rosenbaum J. L. Transcription of alpha- and beta-tubulin genes in vitro in isolated Chlamydomonas reinhardi nuclei. J Cell Biol. 1984 Mar;98(3):1138–1143. doi: 10.1083/jcb.98.3.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kindle K. L. High-frequency nuclear transformation of Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1228–1232. doi: 10.1073/pnas.87.3.1228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kindle K. L., Schnell R. A., Fernández E., Lefebvre P. A. Stable nuclear transformation of Chlamydomonas using the Chlamydomonas gene for nitrate reductase. J Cell Biol. 1989 Dec;109(6 Pt 1):2589–2601. doi: 10.1083/jcb.109.6.2589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knight D. E., Scrutton M. C. Gaining access to the cytosol: the technique and some applications of electropermeabilization. Biochem J. 1986 Mar 15;234(3):497–506. doi: 10.1042/bj2340497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris R. L., Keller L. R., Zweidler A., Rizzo P. J. Analysis of Chlamydomonas reinhardtii histones and chromatin. J Protozool. 1990 Mar-Apr;37(2):117–123. doi: 10.1111/j.1550-7408.1990.tb05880.x. [DOI] [PubMed] [Google Scholar]
- Neumann E., Schaefer-Ridder M., Wang Y., Hofschneider P. H. Gene transfer into mouse lyoma cells by electroporation in high electric fields. EMBO J. 1982;1(7):841–845. doi: 10.1002/j.1460-2075.1982.tb01257.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rochaix J. D., van Dillewijn J. Transformation of the green alga Chlamydomonas reinhardii with yeast DNA. Nature. 1982 Mar 4;296(5852):70–72. doi: 10.1038/296070a0. [DOI] [PubMed] [Google Scholar]
- SAGER R., GRANICK S. Nutritional studies with Chlamydomonas reinhardi. Ann N Y Acad Sci. 1953 Oct 14;56(5):831–838. doi: 10.1111/j.1749-6632.1953.tb30261.x. [DOI] [PubMed] [Google Scholar]
- Schloss J. A., Silflow C. D., Rosenbaum J. L. mRNA abundance changes during flagellar regeneration in Chlamydomonas reinhardtii. Mol Cell Biol. 1984 Mar;4(3):424–434. doi: 10.1128/mcb.4.3.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sokoloski J. A., Jastreboff M. M., Bertino J. R., Sartorelli A. C., Narayanan R. Introduction of deoxyribonucleoside triphosphates into intact cells by electroporation. Anal Biochem. 1986 Nov 1;158(2):272–277. doi: 10.1016/0003-2697(86)90549-x. [DOI] [PubMed] [Google Scholar]
- Widholm J. M. The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 1972 Jul;47(4):189–194. doi: 10.3109/10520297209116483. [DOI] [PubMed] [Google Scholar]
- Zimmermann U., Pilwat G., Riemann F. Preparation of erythrocyte ghosts by dielectric breakdown of the cell membrane. Biochim Biophys Acta. 1975 Jan 28;375(2):209–219. doi: 10.1016/0005-2736(75)90189-3. [DOI] [PubMed] [Google Scholar]



