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Applied and Environmental Microbiology, November 1998, p. 4477-4481, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Sequence Polymorphism in the beta -Tubulin Gene Reveals Heterogeneous and Variable Population Structures in Cryptosporidium parvum

Giovanni Widmer,1,* Laurie Tchack,1 Cynthia L. Chappell,2 and Saul Tzipori1,3

Division of Infectious Diseases, Tufts University School of Veterinary Medicine, North Grafton, Massachusetts 015361; Division of Geographic Medicine, New England Medical Center, Boston, Massachusetts 021113; and University of Texas School of Public Health, Houston, Texas 772252

Received 24 June 1998/Accepted 5 August 1998

Restriction fragment length polymorphism (RFLP) analysis of isolates of Cryptosporidium parvum has revealed two subgroups, termed H and C. The limited resolution of the RFLP method precludes an in-depth study of the genetic structure of C. parvum populations. Published C. parvum restriction polymorphisms lie within protein-coding regions known to be more homogeneous than noncoding sequences. To better assess the degrees of heterogeneity between and within C. parvum isolates, sequence polymorphism in the beta -tubulin intron, the only C. parvum intron described to date, was investigated. In contrast to the two genotypes distinguished by multilocus RFLP, several alleles were detected by sequence and RFLP analysis of the beta -tubulin intron and adjacent exon 2. Isolates carrying different beta -tubulin alleles were found. Significantly, one of the beta -tubulin alleles present in two geographically unrelated isolates combined features of C- and H-type isolates, suggesting that it might have arisen from a recombination event. A comparison of multiple samples of a calf-propagated laboratory isolate showed that the ratio of different beta -tubulin alleles fluctuated during serial passage.

* Corresponding author. Mailing address: Tufts University, Bldg. 20, 200 Westboro Road, North Grafton, MA 01536. Phone: (508) 839 7944. Fax: (508) 839 7977. E-mail: gwidmer{at}infonet.tufts.edu.

Applied and Environmental Microbiology, November 1998, p. 4477-4481, Vol. 64, No. 11
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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