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Plant Sciences

Center for Plant Science Innovation: Faculty and Staff Publications

Series

1997

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Cdna Encoding A Reca Homolog In Eukaryotes, Andre Jagendorf, Heriberto D. Cerutti Oct 1997

Cdna Encoding A Reca Homolog In Eukaryotes, Andre Jagendorf, Heriberto D. Cerutti

Center for Plant Science Innovation: Faculty and Staff Publications

A nucleotide and deduced amino acid sequence of a cDNA encoding the Arabidopsis thulium RecA protein is disclosed.


Epigenetic Silencing Of A Foreign Gene In Nuclear Transformants Of Chlamydomonas, Heriberto D. Cerutti, Anita M. Johnson, Nicholas W. Gillham, John E. Boynton Jun 1997

Epigenetic Silencing Of A Foreign Gene In Nuclear Transformants Of Chlamydomonas, Heriberto D. Cerutti, Anita M. Johnson, Nicholas W. Gillham, John E. Boynton

Center for Plant Science Innovation: Faculty and Staff Publications

The unstable expression of introduced genes poses a serious problem for the application of transgenic technology in plants. In transformants of the unicellular green alga Chlamydomonas reinhardtii, expression of a eubacterial aadA gene, conferring spectinomycin resistance, is transcriptionally suppressed by a reversible epigenetic mechanism(s). Variations in the size and frequency of colonies surviving on different concentrations of spectinomycin as well as the levels of transcriptional activity of the introduced transgene(s) suggest the existence of intermediate expression states in genetically identical cells. Gene silencing does not correlate with methylation of the integrated DNA and does not involve large alterations in its …


A Eubacterial Gene Conferring Spectinomycin Resistance On Chlamydomonas Reinhardtii: Integration Into The Nuclear Genome And Gene Expression, Heriberto D. Cerutti, A. M. Johnson, N. W. Gillham, J. E. Boynton Jan 1997

A Eubacterial Gene Conferring Spectinomycin Resistance On Chlamydomonas Reinhardtii: Integration Into The Nuclear Genome And Gene Expression, Heriberto D. Cerutti, A. M. Johnson, N. W. Gillham, J. E. Boynton

Center for Plant Science Innovation: Faculty and Staff Publications

We have constructed a dominant selectable marker for nuclear transformation of C. reinhardtii, composed of the coding sequence of the eubacterial aadA gene (conferring spectinomycin resistance) fused to the 5' and 3' untranslated regions of the endogenous RbcS2 gene. Spectinomycin-resistant transformants isolated by direct selection (1) contain the chimeric gene(s) stably integrated into the nuclear genome, (2) show cosegregation of the resistance phenotype with the introduced DNA, and (3) synthesize the expected mRNA and protein. Small linearized plasmids appeared to be inserted into the nuclear genome preferentially through their ends, with relatively few large deletions and/or rearrangements. Multiple copy transformants …