2024-03-29T14:44:38Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/2119882020-10-20T12:01:00Zcom_10261_86com_10261_1col_10261_339
DIGITAL.CSIC
author
Ramiro Ibáñez, M. J.
author
Hanke, Tobias
author
Taladriz, Soraya
author
Larraga, Vicente
funder
Ministerio de Ciencia y Tecnología (España)
orcid
Larraga, Vicente [0000-0003-1260-7400]
2020-05-22T12:02:47Z
2020-05-22T12:02:47Z
2002-08
Parasitol Res 88 (8) 760–767 (2002)
0932-0113
http://hdl.handle.net/10261/211988
10.1007/s00436-002-0653-0
1432-1955
http://dx.doi.org/10.13039/501100006280
Gene expression level is extremely difficult to assess in the intracellular form of Leishmania infantum, the amastigote, due to host mRNA contamination, low supply of parasites and stress degradation problems. We obtained and analyzed L. infantum DNA polymerase β (Li Pol β) , a suitable enzyme for differential expression studies. The amount of Li Pol β mRNA was determined in different forms of the parasite (extracellular promastigote, intracellular amastigote) by Northern blot and by relative quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Amastigote transcript levels were determined from both gradient-purified parasites and directly from a population of infected macrophages. The mRNA, undetectable when obtained from amastigotes by the classic gradient centrifugation method and subsequent Northern analysis, was clearly and specifically detectable by this quantitative RT-PCR method from the mixed macrophage/parasite cell population. Li Pol β displays a different pattern of expression in the distinct forms of the parasite cycle that correlate with the infectivity of the protozoon. Thus, Li Pol β mRNA expression is developmentally regulated, being clearly higher in the more infective forms of the parasite.
eng
closedAccess
Infected macrophage
Intracellular form
Intracellular amastigote
Degradation problem
Classic gradient
DNA polymerase beta mRNA determination by relative quantitative RT-PCR from Leishmania infantum intracellular amastigotes
artículo
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URL
https://digital.csic.es/bitstream/10261/211988/1/accesoRestringido.pdf
File
MD5
42637ae8545636bc41605c1740a9a84e
15753
application/pdf
accesoRestringido.pdf