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A Method for Isolation of RNA from Pneumocystis carinii
Author(s) -
CUSHION MELANJE T.,
BLASÉ MARIA AIRO,
VVALZER PETER D.
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.tb02668.x
Subject(s) - ribosomal rna , pneumocystis carinii , biology , rna , isolation (microbiology) , microbiology and biotechnology , biochemistry , virology , human immunodeficiency virus (hiv) , gene , pneumocystis jirovecii
ABSTRACT Toial RNA from Pneumocystis carinii obtained directly from the rat lung and from short term culture on A549 cells was evaluated for size and purity. An isolation procedure using guanidine isothiocyanate and lithium chloride was preferable to a hot phenol method. Host cells were eliminated by hypotonic lysis and a series of microfiltrations. Pneumocystis carinii were pretreated with Zymolyase for increased susceptibility to chaotropic agents. The major ribosomal species of P. carinii RNA migrated similarly to Saccharomyces cerevisiae rRNA. The 28s‐like species migrated well ahead of rat and A549 cell rRNA and weli behind the prokaryotic large rRNA species.