Leishmania (Viannia) guyanensis: A New Minicircle Class Exclusive to This Specie Isolated from a DNA Cosmid Library Useful for Taxonomic Purposes

• Published in: Experimental parasitology 2000-03, Vol.94 (3), p.143-149
• Main Authors: Rodriguez, N., Rodriguez, A., Cardona, M., Barrios, M.A., McCann, S.H.E., Barker, D.C.
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Biological and medical sciences; Blotting, Southern; bp, base pairs; Cloning, Molecular; Conserved Sequence; CSBs, conserved sequence blocks; DNA, Kinetoplast - chemistry; DNA, Protozoan - chemistry; DNA, Recombinant - chemistry; Electrophoresis, Gel, Pulsed-Field; Fundamental and applied biological sciences. Psychology; kb, kilobases; kDNA kinetoplast deoxyribonucleic acid; Leishmania (Viannia) guyanensis; Leishmania guyanensis; Leishmania guyanensis - classification; Leishmania guyanensis - genetics; minicircles; Molecular Sequence Data; PCR, polymerase chain reaction; Polymerase Chain Reaction; Protozoa; Repetitive Sequences, Nucleic Acid; Restriction Mapping; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Systematics. Geographical distribution. Morphology. Cytology; Viannia guyanensis
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1006/expr.1999.4482

Rodriguez, N., Rodriguez, A., Cardona, M., Barrios, M. A., McCann, S. H. E., and Barker, D. C. 2000. Leishmania (Viannia) guyanensis: A new minicircle class exclusive to this specie isolated from a DNA cosmid library useful for taxonomic purposes. Experimental Parasitology94, 143–149. In this paper we describe a new minicircle class exclusive to Leishmania (Viannia) guyanensis. The minicircle class was obtained with the aid of a total DNA cosmid library. The library was screened with an EcoRI fragment isolated from L. (V.) guyanensis (M4147). After screening seven clones were selected which showed strong hybridisation. Clones were digested and hybridised with the same probe. After hybridisation only one clone containing the desired fragment was positive. The fragment sized around 1000 bp was subcloned into pBluescript for sequencing. Sequence analysis using the GCG programme showed no substantial homology with any sequences previously reported, apart from the expected homology with the conserved region of Leishmania kDNA sequences. The probe hybridised strongly only to L. (V.) guyanensis kDNA after medium stringency washing.