Journal of Parasitology

Published by: American Society of Parasitologists

« previous article : next article »


Journal of Parasitology 93(3):724-726. 2007
doi: 10.1645/GE-1099R.1

DNA Probes for Identifying Chromosomes 5, 6, and 7 of Schistosoma mansoni

T. Taguchia, Y. Hiraib, P. T. LoVerdec, A. Tominagaa, and H. Hiraib

aDivision of Human Health and Medical Science, Graduate School of Kuroshio Science, Kochi University, Nankoku, Kochi 783-8505, Japan

bPrimate Research Institute, Kyoto University, Inuyama, Aichi 484-8506, Japan

cSouthwest Foundation for Biomedical Research, San Antonio, Texas 78245.

Schistosoma mansoni has a genome of 270 Mb contained on 8 pairs of chromosomes. C-banding has been a useful technique in identifying the 7 autosomal and sex chromosomes. However, even with C-banding, S. mansoni chromosomes 5, 6, and 7 are difficult to discriminate from each other, because of their small sizes, morphological similarity, and poor banding patterns. We have identified probes that specifically paint chromosomes 5, 6, and 7 of S. mansoni with the use of chromosome microdissection and the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). Exact chromosome identification is required for accurate chromosome mapping of genomic clones and genetic elements, which is an essential component of the schistosome genome project.



LITERATURE CITED

Behrens, F., U. Claussen, L. M. Iyer, E. D. Green, B. Horsthemke, R. Williamson, C. Huxley, and C. Coutelle. 1997. Isolation of DNA from the centromere of human chromosome 7 by microdissection. Chromosome Research 5:215–220. CrossRef, PubMed
Christian, A. T., H. E. Garcia, and J. D. Tucker. 1999. PCR in situ followed by microdissection allows whole chromosome painting probes to be made from single microdissected chromosomes. Mammalian Genome 10:628–631. CrossRef, PubMed, CSA
El-Sayed, N., D. Bartholomeu, A. Ivens, D. A. Johnston, and P. T. LoVerde. 2004. Advances in schistosome genomics. Trends in Parasitology 20:154–157. CrossRef, PubMed
Engels, D., L. Chitsulo, A. Montresor, and L. Savioli. 2002. The global epidemiological situation of schistosomiasis and new approaches to control and research. Acta Tropica 82:139–146. CrossRef, PubMed
Franco, G. R., A. F. Valadao, A. Azevedo, and E. M L. Rabelo. 2000. The Schistosoma gene discovery program: State of the art. International Journal for Parasitology 30:453–463. CrossRef, PubMed, CSA
Hirai, H. and Y. Hirai. 2004. FISH mapping for helminth genome. Methods in Molecular Biology 270:379–394. PubMed
Hirai, H. and P. T. LoVerde. 1995. FISH techniques for constructing physical maps on schistosome chromosomes. Parasitology Today 11:310–314. CrossRef, PubMed, CSA
Hirai, H., T. Taguchi, Y. Saitoh, M. Kawanaka, H. Sugiyama, S. Habe, M. Okamoto, M. Hirata, M. Shimada, W. Tiu, K. Lai, E. S. Upatham, and T. Agatsuma. 2000. Chromosome differentiation of the Schistosoma japonicum complex. International Journal for Parasitology 30:441–452. CrossRef, PubMed, CSA
Le Paslier, M. C., R. J. Pierce, F. Merlin, H. Hirai, W. Wu, D. L. Williams, D. Johnston, P. T. LoVerde, and D. Le Paslier. 2000. Construction and characterization of a Schistosoma mansoni bacterial artificial chromosome library. Genomics 65:87–94. CrossRef, PubMed, CSA
LoVerde, P. T., H. Hirai, J. M. Merrick, N. H. Lee, and N. El-Sayed. 2004. Schistosoma mansoni genome project: An update. Parasitology International 53:183–192. CrossRef, PubMed
Short, R. B. 1983. Presidential address: Sex and the single schistosome. Journal of Parasitology 69:3–22. PubMed
Simpson, A. J G., A. Sher, and T. F. McCutchan. 1982. The genome of Schistosoma mansoni: Isolation of DNA, its size, bases and repetitive sequences. Molecular and Biochemical Parasitology 6:125–137. CrossRef, PubMed
Taguchi, T., K. Akimaru, H. Hirai, Y. Hirai, J. M. Mwenda, and K. Yuri. 2003. A probe generated by chromosome microdissection, useful for analyzing Y chromosome evolution in Old World monkey. Chromosome Research 11:147–152. CrossRef, PubMed
Taguchi, T., G. Z. Cheng, D. W. Bell, B. Balsara, Z. Liu, J. M. Siegfried, and J. R. Testa. 1997. Combined chromosome microdissection and comparative genomic hybridization detect multiple sites of amplification DNA in a human lung carcinoma cell line. Genes, Chromosomes & Cancer 20:208–212. CrossRef, PubMed, CSA
Taguchi, T., H. Hirai, E. S. Upatham, and K. Agatsuma. 2000. Sex chromosome differences among the three races (alpha, beta, gamma) of the snail intermediate host of Schistosoma mekongi, Neotricula aperta. Parasitology International 49:267–272. CrossRef, PubMed, CSA
Tanaka, M., H. Hirai, P. T. LoVerde, S. Nagafuchi, G. R. Franco, A. J. Simpson, and S. D. Pena. 1995. Yeast artificial chromosome (YAC)-based genome mapping of Schistosoma mansoni. Molecular and Biochemical Parasitology 69:41–51. CrossRef, PubMed, CSA

Figure 1.A 3-color FISH image of metaphase chromosomes of Schistosoma mansoni painted by microdissection probes obtained from chromosomes 5, 6, and 7. Left and right panels show (a) DAPI-stained and (b) FISH images, respectively. Green FISH signals identify chromosome 5, pink for chromosome 6, and yellow for chromosome 7. Note that chromosomes 5, 6, and 7 are easily distinguished from the other chromosomes in the complement by the specific chromosome color. Scale is 10 μm

click this button to close

Article Views

Article Tools

click this button to open

Citing Articles

 
BioOne is the product of innovative collaboration between scientific societies, libraries, academe and the private sector.
 
21 Dupont Circle NW, Suite 800, Washington, DC 20036 • Phone 202.296.2296 • Fax 202.872.0884
 
Copyright © 2009 BioOne All rights reserved