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. 1995 Mar;7(3):287–294. doi: 10.1105/tpc.7.3.287

Site-selected transposon mutagenesis at the hcf106 locus in maize.

L Das 1, R Martienssen 1
PMCID: PMC160782  PMID: 7734963

Abstract

The High chlorophyll fluorescence106 (Hcf106) gene in maize is required for chloroplast membrane biogenesis, and the hcf106-mum1 allele is caused by the insertion of a Robertson's Mutator Mu1 element into the promoter of the gene. Seedlings homozygous for hcf106-mum1 are pale green and die 3 weeks after germination, but only in the presence of Mutator activity conferred by active, autonomous Mu regulatory transposons elsewhere in the genome. When Mutator activity is lost, the mutant phenotype is suppressed, and homozygous plants have an almost wild-type phenotype. To isolate derivative alleles at the hcf106 locus that no longer require Mutator activity for phenotypic expression, we have developed a method for site-selected transposon mutagenesis in maize. This procedure, first described for Caenorhabditis elegans and Drosophila, involves using polymerase chain reaction (PCR) to screen pools of individuals for insertions and deletions in genes of known sequence. Pools of seedlings segregating for the progenitor allele hcf106-mum1 were screened by PCR for insertions and deletions associated with Robertson's Mutator. In a 360-bp target region, two new insertions and one deletion were identified in only 700 Mu-active gametes screened. One of the insertions was in the progenitor hcf106-mum1 allele and the other was in the wild-type allele, but all three new alleles were found to have break-points at the same nucleotide in the first intron. Unlike the hcf-106-mum1 progenitor allele, the deletion and one of the insertions conferred pale green seedling lethal phenotypes in the absence of mutator activity. However, the second insertion had a weak, viable phenotype under these conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Ballinger D. G., Benzer S. Targeted gene mutations in Drosophila. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9402–9406. doi: 10.1073/pnas.86.23.9402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barkan A., Martienssen R. A. Inactivation of maize transposon Mu suppresses a mutant phenotype by activating an outward-reading promoter near the end of Mu1. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3502–3506. doi: 10.1073/pnas.88.8.3502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barkan A., Miles D., Taylor W. C. Chloroplast gene expression in nuclear, photosynthetic mutants of maize. EMBO J. 1986 Jul;5(7):1421–1427. doi: 10.1002/j.1460-2075.1986.tb04378.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bingham P. M., Judd B. H. A copy of the copia transposable element is very tightly linked to the Wa allele at the white locus of D. melanogaster. Cell. 1981 Sep;25(3):705–711. doi: 10.1016/0092-8674(81)90177-x. [DOI] [PubMed] [Google Scholar]
  5. Chandler V. L., Hardeman K. J. The Mu elements of Zea mays. Adv Genet. 1992;30:77–122. doi: 10.1016/s0065-2660(08)60319-3. [DOI] [PubMed] [Google Scholar]
  6. Chandler V. L., Walbot V. DNA modification of a maize transposable element correlates with loss of activity. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1767–1771. doi: 10.1073/pnas.83.6.1767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fedoroff N. V., Furtek D. B., Nelson O. E. Cloning of the bronze locus in maize by a simple and generalizable procedure using the transposable controlling element Activator (Ac). Proc Natl Acad Sci U S A. 1984 Jun;81(12):3825–3829. doi: 10.1073/pnas.81.12.3825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Greenstein D., Hird S., Plasterk R. H., Andachi Y., Kohara Y., Wang B., Finney M., Ruvkun G. Targeted mutations in the Caenorhabditis elegans POU homeo box gene ceh-18 cause defects in oocyte cell cycle arrest, gonad migration, and epidermal differentiation. Genes Dev. 1994 Aug 15;8(16):1935–1948. doi: 10.1101/gad.8.16.1935. [DOI] [PubMed] [Google Scholar]
  9. Hake S. Unraveling the knots in plant development. Trends Genet. 1992 Mar;8(3):109–114. doi: 10.1016/0168-9525(92)90199-e. [DOI] [PubMed] [Google Scholar]
  10. Hershberger R. J., Warren C. A., Walbot V. Mutator activity in maize correlates with the presence and expression of the Mu transposable element Mu9. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10198–10202. doi: 10.1073/pnas.88.22.10198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaiser K., Goodwin S. F. "Site-selected" transposon mutagenesis of Drosophila. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1686–1690. doi: 10.1073/pnas.87.5.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Martienssen R., Barkan A., Taylor W. C., Freeling M. Somatically heritable switches in the DNA modification of Mu transposable elements monitored with a suppressible mutant in maize. Genes Dev. 1990 Mar;4(3):331–343. doi: 10.1101/gad.4.3.331. [DOI] [PubMed] [Google Scholar]
  13. Martienssen R., Baron A. Coordinate suppression of mutations caused by Robertson's mutator transposons in maize. Genetics. 1994 Mar;136(3):1157–1170. doi: 10.1093/genetics/136.3.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McCLINTOCK B. The origin and behavior of mutable loci in maize. Proc Natl Acad Sci U S A. 1950 Jun;36(6):344–355. doi: 10.1073/pnas.36.6.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ortiz D. F., Strommer J. N. The Mu1 maize transposable element induces tissue-specific aberrant splicing and polyadenylation in two Adh1 mutants. Mol Cell Biol. 1990 May;10(5):2090–2095. doi: 10.1128/mcb.10.5.2090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Qin M. M., Robertson D. S., Ellingboe A. H. Cloning of the Mutator transposable element MuA2, a putative regulator of somatic mutability of the a1-Mum2 allele in maize. Genetics. 1991 Nov;129(3):845–854. doi: 10.1093/genetics/129.3.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rushforth A. M., Saari B., Anderson P. Site-selected insertion of the transposon Tc1 into a Caenorhabditis elegans myosin light chain gene. Mol Cell Biol. 1993 Feb;13(2):902–910. doi: 10.1128/mcb.13.2.902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Zwaal R. R., Broeks A., van Meurs J., Groenen J. T., Plasterk R. H. Target-selected gene inactivation in Caenorhabditis elegans by using a frozen transposon insertion mutant bank. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7431–7435. doi: 10.1073/pnas.90.16.7431. [DOI] [PMC free article] [PubMed] [Google Scholar]

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