Monitoring the Transcriptional Activity of Human Endogenous Retroviral HERV-W Family Using PNA Strand Invasion into Double-Stranded DNA.
Mol Biotechnol
; 60(2): 124-133, 2018 Feb.
Article
en En
| MEDLINE
| ID: mdl-29313202
ABSTRACT
In the presented assay, we elaborated a method for distinguishing sequences that are genetically closely related to each other. This is particularly important in a situation where a fine balance of the allele abundance is a point of research interest. We developed a peptide nucleic acid (PNA) strand invasion technique for the differentiation between multiple sclerosis-associated retrovirus (MSRV) and ERVWE1 sequences, both molecularly similar, belonging to the human endogenous retrovirus HERV-W family. We have found that this method may support the PCR technique in screening for minor alleles which, in certain conditions, may be undetected by the standard PCR technique. We performed the analysis of different ERVWE1 and MSRV template mixtures ranging from 0 to 100% of ERVWE1 in the studied samples, finding the linear correlation between template composition and signal intensity of final reaction products. Using the PNA strand invasion assay, we were able to estimate the relative ERVWE1 expression level in human specimens such as U-87 MG, normal human astrocytes cell lines and placental tissue. The results remained in concordance with those obtained by semi-quantitative or quantitative PCR.
Palabras clave
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Placenta
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Proteínas Gestacionales
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Replicación Viral
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Productos del Gen env
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Astrocitos
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Retrovirus Endógenos
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Ácidos Nucleicos de Péptidos
Límite:
Adult
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Female
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Humans
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Pregnancy
Idioma:
En
Revista:
Mol Biotechnol
Asunto de la revista:
BIOLOGIA MOLECULAR
/
BIOTECNOLOGIA
Año:
2018
Tipo del documento:
Article
País de afiliación:
Polonia