Characterization and partial nucleotide sequence of the DNA fingerprinting probe Ca3 of Candida albicans

J. Anderson, Srikantha Thyagarajan, B. Morrow, S. H. Miyasaki, T. C. White, N. Agabian, J. Schmid, D. R. Soll

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The moderately repetitive Ca3 fragment of Candida albicans has been used as an effective DNA fingerprinting probe in epidemiological studies. EcoRI digestion of Ca3 DNA results in seven fragments of 4.2 kb (A), 2.98 kb (B), 2.85 kb (C), 0.77 kb (D1), 0.77 kb (D2), 0.38 kb (E), and 0.30 kb (F). Five of these EcoRI fragments have been mapped in the 5'-3' order C B D1 A D2. The intact Ca3 probe and the three largest EcoRI fragments, A, B, and C, were individually used to probe Southern blots of EcoRI-digested DNA of a set of test strains, transverse alternating field electrophoresis-separated chromosomes of strain 3153A, and Northern (RNA) blots of test strain 3153A. Fragments A, B, and C each generate a different Southern blot hybridization pattern with EcoRI-digested whole-cell DNA; Ca3 sequences are present in at least five of seven separable chromosomes and a minichromosome of strain 3153A; fragments A, B, and C are distributed differently on chromosomes; and fragments A, B, and C do not cross-hybridize. Ca3 hybridizes to three major transcripts of 2.8, 2.3, and 1.5 kb. Fragment A hybridizes intensely to the 1.5-kb transcript, while fragments B and C both hybridize intensely to the 2.8- and 2.3-kb transcripts. The B fragment, which contains 2,980 bp and contributes to the major portion of the Ca3 pattern, was sequenced. Both direct and inverted repeat sequence motifs were identified. These results provide us with initial insights into the evolution of the Ca3 pattern and the nature of the probe.

Original languageEnglish
Pages (from-to)1472-1480
Number of pages9
JournalJournal of Clinical Microbiology
Volume31
Issue number6
StatePublished - 1 Jan 1993

Fingerprint

DNA Fingerprinting
DNA Probes
Candida albicans
Chromosomes
Southern Blotting
Inverted Repeat Sequences
Nucleic Acid Repetitive Sequences
DNA
Northern Blotting
Electrophoresis
Epidemiologic Studies
Digestion
RNA

Cite this

Anderson, J., Thyagarajan, S., Morrow, B., Miyasaki, S. H., White, T. C., Agabian, N., ... Soll, D. R. (1993). Characterization and partial nucleotide sequence of the DNA fingerprinting probe Ca3 of Candida albicans. Journal of Clinical Microbiology, 31(6), 1472-1480.
Anderson, J. ; Thyagarajan, Srikantha ; Morrow, B. ; Miyasaki, S. H. ; White, T. C. ; Agabian, N. ; Schmid, J. ; Soll, D. R. / Characterization and partial nucleotide sequence of the DNA fingerprinting probe Ca3 of Candida albicans. In: Journal of Clinical Microbiology. 1993 ; Vol. 31, No. 6. pp. 1472-1480.
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Anderson, J, Thyagarajan, S, Morrow, B, Miyasaki, SH, White, TC, Agabian, N, Schmid, J & Soll, DR 1993, 'Characterization and partial nucleotide sequence of the DNA fingerprinting probe Ca3 of Candida albicans', Journal of Clinical Microbiology, vol. 31, no. 6, pp. 1472-1480.

Characterization and partial nucleotide sequence of the DNA fingerprinting probe Ca3 of Candida albicans. / Anderson, J.; Thyagarajan, Srikantha; Morrow, B.; Miyasaki, S. H.; White, T. C.; Agabian, N.; Schmid, J.; Soll, D. R.

In: Journal of Clinical Microbiology, Vol. 31, No. 6, 01.01.1993, p. 1472-1480.

Research output: Contribution to journalArticle

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AU - Anderson, J.

AU - Thyagarajan, Srikantha

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N2 - The moderately repetitive Ca3 fragment of Candida albicans has been used as an effective DNA fingerprinting probe in epidemiological studies. EcoRI digestion of Ca3 DNA results in seven fragments of 4.2 kb (A), 2.98 kb (B), 2.85 kb (C), 0.77 kb (D1), 0.77 kb (D2), 0.38 kb (E), and 0.30 kb (F). Five of these EcoRI fragments have been mapped in the 5'-3' order C B D1 A D2. The intact Ca3 probe and the three largest EcoRI fragments, A, B, and C, were individually used to probe Southern blots of EcoRI-digested DNA of a set of test strains, transverse alternating field electrophoresis-separated chromosomes of strain 3153A, and Northern (RNA) blots of test strain 3153A. Fragments A, B, and C each generate a different Southern blot hybridization pattern with EcoRI-digested whole-cell DNA; Ca3 sequences are present in at least five of seven separable chromosomes and a minichromosome of strain 3153A; fragments A, B, and C are distributed differently on chromosomes; and fragments A, B, and C do not cross-hybridize. Ca3 hybridizes to three major transcripts of 2.8, 2.3, and 1.5 kb. Fragment A hybridizes intensely to the 1.5-kb transcript, while fragments B and C both hybridize intensely to the 2.8- and 2.3-kb transcripts. The B fragment, which contains 2,980 bp and contributes to the major portion of the Ca3 pattern, was sequenced. Both direct and inverted repeat sequence motifs were identified. These results provide us with initial insights into the evolution of the Ca3 pattern and the nature of the probe.

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