Document 0742
 DOCN  M9550742
 TI    Inhibition of HIV type 1 reverse transcriptase assay by nucleases
       produced by contaminating mycoplasmas.
 DT    9505
 AU    Quillent C; Grau O; Clavel F; Montagnier L; Blanchard A; Departement du
       SIDA et des Retrovirus, Institut Pasteur, U.R.A.; CNRS 1157, Paris,
       France.
 SO    AIDS Res Hum Retroviruses. 1994 Oct;10(10):1251-7. Unique Identifier :
       AIDSLINE MED/95151362
 AB    Mycoplasmal contamination of HIV-1-infected cells has been found to
       induce reduction of reverse transcriptase (RT) activity; however, the
       exact mechanism of this phenomenon was not clearly elucidated. Our
       results indicate that the apparent reduction in RT activity is due to a
       calcium-dependent nuclease(s) that is (are) produced by contaminating
       mycoplasmas. The interference with the RT assay was found to be due to
       the degradation of products of the RT activity. Addition of EGTA at a 1
       mM concentration was sufficient to remove the inhibitory effect. The
       particular HIV-1-producing cell line that was under study was found to
       be contaminated with Mycoplasma fermentans and Mycoplasma pirum and the
       latter was isolated in pure culture. Nuclease activity was also observed
       with pure cultures of mycoplasmas from different species. The activity
       was found to be of the endonuclease type because it was active with both
       supercoiled and linear DNAs.
 DE    Cell Line  Deoxyribonucleases/BIOSYNTHESIS/*PHARMACOLOGY  DNA Primers
       DNA, Bacterial/ANALYSIS  Equipment Contamination  Human
       HIV-1/*ENZYMOLOGY  Mycoplasma/DRUG EFFECTS/ENZYMOLOGY/*ISOLATION & PURIF
       Mycoplasma fermentans/ISOLATION & PURIF  Polymerase Chain
       Reaction/METHODS  Quinolones/PHARMACOLOGY  Reverse
       Transcriptase/*ANTAGONISTS & INHIB/ANALYSIS  Substrate Specificity
       Support, Non-U.S. Gov't  T-Lymphocytes  Tissue Culture  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
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