|
|
|
|
|
| Zhao,Yongliang; Dong,Zongming; Li,Tengfei; Huang,Gang. |
| Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection... |
| Tipo: Journal article |
Palavras-chave: Adenovirus; AdEasy; Colony PCR; Homologous recombination; RecA. |
| Ano: 2014 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008 |
| |
|
|
| Wang,Jianmin; Song,Ruihua; Chen,Lei; Yin,Liangjun; Chen,Bo; Sun,Jing; Gou,Yuanbing; Zhao,Ling; Chen,Lin. |
| Objective: To investigate the functions of Fibroblast Growth Factor Receptor-2 (FGFR2) at different stages of cell differentiation. The engineered murine embryonic stem (ES) cells with conditional knockout of FGFR2 were developed depending on Cre-loxP. Methods: Cre-loxP system was used in a conditional targeting vector. The competent AM-1 bacteria, which expressed Cre-recombinase, was used to confirm the Cre-mediated deletion of the floxed exons 7 and 8 of FGFR2. The targeting vector was electroporated into the ES cells, and the transfected ES cells were screened with G418 and Ganciclovir. Finally, the ES clones with correct targeting events were identified by Southern Blot and PCR. Results: The targeting vector with conditional knockout of murine FGFR2... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Fibroblast growth factor receptor-2 (FGFR2); Conditional knockout; Homologous recombination; Cre; ES cells. |
| Ano: 2006 |
URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452006000200002 |
| |
|
| |
|
|
| ROCHA-MARTINS,MAURÍCIO; CAVALHEIRO,GABRIEL R.; MATOS-RODRIGUES,GABRIEL E.; MARTINS,RODRIGO A.P.. |
| ABSTRACTGenome modification technologies are powerful tools for molecular biology and related areas. Advances in animal transgenesis and genome editing technologies during the past three decades allowed systematic interrogation of gene function that can help model how the genome influences cellular physiology. Genetic engineering via homologous recombination (HR) has been the standard method to modify genomic sequences. Nevertheless, nuclease-guided genome editing methods that were developed recently, such as ZFN, TALEN and CRISPR/Cas, opened new perspectives for biomedical research. Here, we present a brief historical perspective of genome modification methods, focusing on transgenic mice models. Moreover, we describe how new techniques were discovered... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Homologous recombination; DNA repair; Cre-LoxP; ZFN; TALEN; CRISPR/Cas9.. |
| Ano: 2015 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652015000301323 |
| |
|
|
| ESQUISSATO,GIOVANA N.M.; SANT'ANNA,JULIANE R. DE; FRANCO,CLAUDINÉIA C.S.; ROSADA,LÚCIA J.; SANTOS,PAULA A.S.R. DOS; CASTRO-PRADO,MARIALBA A.A. DE. |
| Mitotic recombination is a process involved in carcinogenesis which can lead to genetic loss through the loss of heterozygosity. The recombinogenic potentials of two anticancer drugs topoisomerase I inhibitors, camptothecin (CPT) and irinotecan (CPT-11), were evaluated in the present study. The homozygotization assay, which assess the induction of mitotic recombination and gene homozygosis, as well as the heterozygous A757//UT448 diploid strain of Aspergillus nidulanswere employed. The three non-cytotoxic concentrations of CPT (3.5 ng mL−1, 10.5 ng mL−1 and 17.4 ng mL−1) were found to induce both mitotic recombination and gene homozygosis. CPT treatment produced three diploids homozygous, for nutritional and conidia color genes, and Homozygotization... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Anticancer drugs; Homozygotization assay; Homologous recombination; Secondary malignancies. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652014000401703 |
| |
|
|
| Paviolo,Natalia Soledad; Vega,María Belén de la; Pansa,María Florencia; García,Iris Alejandra; Calzetta,Nicolás Luis; Soria,Gastón; Gottifredi,Vanesa. |
| Abstract The poly (adenosine diphosphate (ADP)-ribosyl) polymerase inhibitors (PARPi) selectively kill cancer cells with BRCA1 or BRCA2 (BRCA)-mutations. It has been proposed that cell death induction after PARPi depends on unrepaired double strand breaks (DSBs) that accumulate due to the homologous recombination deficiency of BRCA-mutated cells. Such accumulation of DSBs is inferred mainly from the high levels of DNA damage markers like phosphorylated histone H2AX. Herein, we developed a model of isogenic cell lines to show that depletion of BRCA causes PARPi-triggered cell death, replication stress (phosphorylated-H2AX and 53BP1 foci), and genomic instability. However, persistent DSBs accumulation was not detected under the same experimental conditions.... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: GammaH2AX; Alternative end joining; Non-homologous end joining; Homologous recombination; PARP. |
| Ano: 2020 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572020000200303 |
| |
|
| |
|
|
| Hogrel, Gaelle; Lu, Yang; Alexandre, Nicolas; Bossé, Audrey; Dulermo, Remi; Ishino, Sonoko; Ishino, Yoshizumi; Flament, Didier. |
| Among the three domains of life, the process of homologous recombination (HR) plays a central role in the repair of double-strand DNA breaks and the restart of stalled replication forks. Curiously, main protein actors involved in the HR process appear to be essential for hyperthermophilic Archaea raising interesting questions about the role of HR in replication and repair strategies of those Archaea living in extreme conditions. One key actor of this process is the recombinase RadA, which allows the homologous strand search and provides a DNA substrate required for following DNA synthesis and restoring genetic information. DNA polymerase operation after the strand exchange step is unclear in Archaea. Working with Pyrococcus abyssi proteins, here we show... |
| Tipo: Text |
Palavras-chave: Homologous recombination; Archaea; DNA polymerase; Recombinase; DNA repair. |
| Ano: 2020 |
URL: https://archimer.ifremer.fr/doc/00641/75274/75539.pdf |
| |
|
|
|
