Use of a cysteine proteinase from Carica candamarcensis as a protective agent during DNA extraction.
We describe the use of a plant cysteine proteinase isolated from latex of Carica candamarcensis as a protective agent during isolation of bacterial DNA following growth in culture of these cells. Between 100 to 720 units of proteinase (1 microgram = 6 units) afforded good DNA protection when incubated with various kinds of microorganisms. Agarose gel electrophoresis showed that the resulting DNA was similar in size to DNA preparations obtained by treatment with proteinase K. The viability of the resulting material was checked by PCR amplification using species-specific primers. After standing at room temperature (25 degrees C) for 35 days, the enzyme lost 10% of its initial activity. The enzyme stability and good yield of DNA suggest the use of this proteinase as an alternative to proteinase K.
Genelhu
M S
MS
Departamento de Bioquímica, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil.
Zanini
M S
MS
Veloso
I F
IF
Carneiro
A M
AM
Lopes
M T
MT
Salas
C E
CE
D000818:Animals; D003546:Cysteine Endopeptidases / Q000494:pharmacology*; D004247:DNA / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D019286:Endopeptidase K / Q000494:pharmacology; D006023:Glycoproteins / Q000494:pharmacology*; D007919:Leptospira; D016133:Polymerase Chain Reaction; D020011:Protective Agents / Q000494:pharmacology*
D004269:DNA, Bacterial; D006023:Glycoproteins; D020011:Protective Agents; D004247:DNA; D019286:Endopeptidase K; C090775:Carica candamarencis glycosylated cysteine proteinase; D003546:Cysteine Endopeptidases