Construction of an adenoviral vector carrying the CDNA of XPD gene for complementation of human cells deficient in DNA repair (2002)
- Autor:
- Autor USP: MENCK, CARLOS FREDERICO MARTINS - ICB
- Unidade: ICB
- Assunto: MICROBIOLOGIA
- Language: Inglês
- Abstract: In all living organisms, DNA is continually exposed to agents that cause damage to its structure. NER (Nucleotide Excision Repair) is the most flexible of all known DNA repair mechanisms. XPD is one of the proteins involved in NER. It is a subunit of TFIIH complex and exhibits helicase function. Different mutations in XPD can cause different genetic disorders as Xeroderma Pigmentosum group-D (XP-D), XP combined with Cockayne Syndrome (XPD/CS) and Trichothiodystrofy (TTD). Among these syndromes, only XP-D patients have skin cancers. All of them have mental retardation and their cells are sensitive to killing by UV ligth. Fibroblasts from patients of these syndromes showed lower levels of Unscheduled DNA Synthesis (UDS) than normal ones. For complementation studies of these cells, we have constructed a recombinant adenovirus (AdSHIRES-XPD), deleted in E1 and E3 regions, carrying the cDNA of XPD and the EGFP reporter gene linked by an IRES sequence (Internal Ribosome Entry Site). This construction permits both the gene of interest (cloned into the MCS) and the EGFP gene to be translated from a single bicistronic mRNA. The cassette containing the IRES-EGFP-XPD was cloned into the adenoviral vector (pAdeno-X). The recombinant adenoviral construct was cleaved with Pac I to expose its inverted terminal repeats, and transfected into human HEK 293 packaging cells. The process of viral production could be followed in the HEK 293 cells by visualization of the EGFPprotein in a fluorescence microscope. Cytopathic effect appears 10 days after transfection and the resultant virus is now being checked for the presence of XPD cDNA. The characterized virus will be used for infection in XPD, XPD/CS and TTD primary fibroblasts cells, obtained from patients biopses. The virus efficiency will be evaluated by cell survival and recovery by UDS activity
- Imprenta:
- Publisher: Comissão de Cultura e Extensão Universitária do ICB/USP
- Publisher place: São Paulo
- Date published: 2002
- Source:
- Título: Resumos
- Conference titles: Congresso Instituto Ciências Biomédicas, IV
-
ABNT
MENCK, Carlos Frederico Martins. Construction of an adenoviral vector carrying the CDNA of XPD gene for complementation of human cells deficient in DNA repair. 2002, Anais.. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP, 2002. . Acesso em: 29 dez. 2025. -
APA
Menck, C. F. M. (2002). Construction of an adenoviral vector carrying the CDNA of XPD gene for complementation of human cells deficient in DNA repair. In Resumos. São Paulo: Comissão de Cultura e Extensão Universitária do ICB/USP. -
NLM
Menck CFM. Construction of an adenoviral vector carrying the CDNA of XPD gene for complementation of human cells deficient in DNA repair. Resumos. 2002 ;[citado 2025 dez. 29 ] -
Vancouver
Menck CFM. Construction of an adenoviral vector carrying the CDNA of XPD gene for complementation of human cells deficient in DNA repair. Resumos. 2002 ;[citado 2025 dez. 29 ] - DNA Polimerase in sugar cane Xylella fastidiosa and Xanthomonas axonopodis pv. citri
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