Abstract
A sensitive in situ hybridization technique which was effective for mapping genes of low copy number on human metaphase chromosomes was used for gene mapping on maize pachytene chromosomes. A cloned genomic EcoR1 fragment of 10.8 kb, containing most or all of the sequence encoding the Waxy locus mRNA, was used as the probe. Southern DNA blotting analyses performed by Shure et al. (1983) indicated that the Waxy locus was a single copy sequence. In our in situ hybridization experiment, the probe hybridized to a specific site on chromosome 9. Labeling at this site was detected in 48.6% of 154 randomly selected copies of chromosome 9. To test the sensitivity of the method, subclones of the fragment with insert sizes of 6.6, 4.7, 3.5, 2.3, 1.9 and 0.8 kb were used for in situ hybridizations. Labeling efficiency for each probe was determined. The data showed that a single copy probe of 1.9 kb could be detected at the correct position in 18% of 183 randomly selected number 9 chromosomes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ambros PF, Matzke MA, Matzke AJM (1986) Detection of a 17 kb unique sequence (T-DNA) in plant chromosomes by in situ hybridization. Chromosoma 94:11–18
Coe EH Jr, Neuffer MG (1979) The genetics of corn. In: Sprague GF (ed) Corn and corn improvement, 2nd ed. Agronomy 18, Am Soc Agron, Madison, Wis, pp 111–223
Gerhard DA, Kawasaki ES, Bancroft FC, Szabo P (1981) Localization of a unique gene by direct hybridization in situ. Proc Natl Acad Sci USA 78:3755–3759
Greilhuber J (1979) Why plant chromosomes do not show G-bands. Theor Appl Genet 50:121–124
Harper ME, Saunders GF (1981) Localization of single copy DNA sequences on G-banded human chromosomes by in situ hybridization. Chromosoma 83:431–439
Jhanwar SC, Neel BG, Hayward WS, Changanti RSK (1983) Localization of c-ras oncogene family on human germ-line chromosomes. Proc Natl Acad Sci USA 80:4794–4797
Landegent JE, Jansen in de Wal N, van Ommen G-JB, Baas F, de Vijlder JJM, van Duijn P, van der Ploeg M (1985) Chromosomal localization of a unique gene by non-autoradiographic in situ hybridization. Nature 317:175–177
Leary JJ, Brigati DJ, Ward DC (1983) Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bioblots. Proc Natl Acad Sci USA 80:4045–4049
Li C, Wu M, Margitich IS, Davidson N (1986) Gene mapping on human metaphase chromosomes by in situ hybridization with 3H, 35S and 32p labeled probes and transmission electron microscopy. Chromosoma 93:305–312
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press, New York, pp 86–87
Mascia PN, Rubenstein I, Phillips RL, Wang AS, Lu ZX (1981) Localization of the 5s rRNA genes and evidence for diversity in the 5S rDNA region of maize. Gene 15:7–20
Neuffer MG, Jones L, Zuber MS (1968) Mutants of maize. Crop Sci Soc Am, Madison, Wis, p 4
Phillips RL, Wang AS (1982) In situ hybridization with meiotic cells. In: Sheridan WF (ed) Maize for biological research. Plant Mol Biol Ass, Va, pp 121–122
Shen D, Wu M (1987) Transmission electron microscopic study of maize pachytene chromosome 6 Chromosoma
Shure M, Wessler S, Federoff N (1983) Molecular identification and isolation of the Waxy locus in maize. Cell 35:225–233
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shen, Dl., Wang, Zf. & Wu, M. Gene mapping on maize pachytene chromosomes by in situ hybridization. Chromosoma 95, 311–314 (1987). https://doi.org/10.1007/BF00293177
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00293177