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Characterization and Cloning of Pneumocystis carinii Nucleic Acid
Author(s) -
WORLEY MARK A.,
IVEY MICHAEL H.,
GRAVES DON C.
Publication year - 1989
Publication title -
the journal of protozoology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 0022-3921
DOI - 10.1111/j.1550-7408.1989.tb05809.x
Subject(s) - pneumocystis carinii , nucleic acid , microbiology and biotechnology , rna , dna , biology , proteinase k , ribosomal rna , virology , biochemistry , gene , human immunodeficiency virus (hiv) , pneumocystis jirovecii
.Large numbers of Pneumocystis carinii (2 × 10 10 nuclei) were isolated and separated from the lungs of immunosuppressed rats by an enzymatic (collagenase, hyaluronidase and DNase) digestion procedure. The nucleic acid isolated from this P. carinnii‐enriched preparation was characterized by melting point analysis and RNA‐sizing gels. The GC content of P. carinii DNA was approximately 33% while the rat DNA was 41.4%. In addition, RNA isolated from the P. carinii‐enriched preparation showed unique ribosomal RNA bands of 3.4 kb and 1.8 kb as compared with uninfected rat lung ribosomal RNA which banded at 4.8 and 1.9 kb. Following isolation and fragmentation by sonication, the P. carinii DNA fragments were inserted into the vector, λ gt‐11. The resultant library contained 1.1 × 10 5 phage, of which 40–45% hybridized to P. carinii DNA but not to rat DNA.