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An electrophoretic karyotype and assignment of ribosomal genes to resolved chromosomes of Pneumocystis carinii
Author(s) -
Yoganathan T.,
Lin H.,
Buck G. A.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb00132.x
Subject(s) - pneumocystis carinii , biology , karyotype , chromosome , microbiology and biotechnology , ribosomal rna , genetics , gel electrophoresis , homologous chromosome , genome , gene , southern blot , agarose gel electrophoresis , pulsed field gel electrophoresis , virology , human immunodeficiency virus (hiv) , genotype , pneumocystis jirovecii
Summary Pulsed‐field gel electrophoresis was used to generate a molecular karyotype of chromosomes from the opportunistic AIDS pathogen, Pneumocystis carinii. P. carinii cysts and trophozoites were isolated from immunosuppressed rats, lysed in situ in agarose blocks, and subjected to orthogonal‐field gel electrophoresis (OFAGE) and contour‐clamped homogeneous‐field gel electrophoresis (CHEF). OFAGE and CHEF gels resolved, respectively, 16 or 20 chromosome bands ranging in size from 0.32‐1.5 megabase pairs. Summation of the estimated sizes of these chromosomes suggested a total genome complexity for P. carinii of 8‐16 megabase pairs. Homologous probes for the genes encoding the 18S, 5.8S, and 5S ribosomal RNAs were hybridized to filter blots of the pulsed‐field gels to map these genes to specific P. carinii chromosomes.