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| Wang,Wenwen; Liu,Hongmao; Zhao,Haibin; Luo,Zaili; Shi,Yongxiang. |
| This work studied the design, construction, and cleavage analysis of zinc finger nucleases (ZFNs) that could cut the specific sequences within microphthalmia - associate transcription factor (mitfa) of zebra fish. The target site and ZFPs were selected and designed with zinc finger tools, while the ZFPs were synthesized using DNAWorks and two-step PCR. The ZFNs were constructed, expressed, purified, and analyzed in vitro. As expected, the designed ZFNs could create a double-stand break (DSB) at the target site in vitro. The DNAWorks, two-step PCR, and an optimized process of protein expression were firstly induced in the construction of ZFNs successfully, which was an effective and simplified protocol. These results could be useful for further application... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Zinc finger nuclease; DNAWorks; Gene synthesis; Gene knockout. |
| Ano: 2012 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132012000400011 |
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| Huang,P.L.. |
| Targeted disruption of the neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) genes has led to knockout mice that lack these isoforms. These animal models have been useful to study the roles of nitric oxide (NO) in physiologic processes. nNOS knockout mice have enlarged stomachs and defects in the inhibitory junction potential involved in gastrointestinal motility. eNOS knockout mice are hypertensive and lack endothelium-derived relaxing factor activity. When these animals are subjected to models of focal ischemia, the nNOS mutant mice develop smaller infarcts, consistent with a role for nNOS in neurotoxicity following cerebral ischemia. In contrast, eNOS mutant mice develop larger infarcts, and show a more pronounced... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Nitric oxide; Targeted disruption; Gene knockout; Endothelium; Stroke; Atherosclerosis. |
| Ano: 1999 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999001100005 |
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| Zhang,Dalong; Guan,Dan; Liang,Jingbo; Guo,Chunqian; Xie,Xixian; Zhang,Chenglin; Xu,Qingyang; Chen,Ning. |
| L-lactate is one of main byproducts excreted in to the fermentation medium. To improve L-glutamate production and reduce L-lactate accumulation, L-lactate dehydrogenase-encoding gene ldhA was knocked out from L-glutamate producing strain Corynebacterium glutamicum GDK-9, designated GDK-9ΔldhA. GDK-9ΔldhA produced approximately 10.1% more L-glutamate than the GDK-9, and yielded lower levels of such by-products as α-ketoglutarate, L-lactate and L-alanine. Since dissolved oxygen (DO) is one of main factors affecting L-lactate formation during L-glutamate fermentation, we investigated the effect of ldhA deletion from GDK-9 under different DO conditions. Under both oxygen-deficient and high oxygen conditions, L-glutamate production by GDK-9ΔldhA was not higher... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Corynebacterium glutamicum; L-glutamate; L-lactate dehydrogenase; Gene knockout; Fermentation. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000400044 |
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| Nerys-Junior,Arildo; Costa,Lendel C.; Braga-Dias,Luciene P.; Oliveira,Márcia; Rossi,Átila D.; Cunha,Rodrigo Delvecchio da; Gonçalves,Gabriel S.; Tanuri,Amilcar. |
| Engineered nucleases such as zinc finger nucleases (ZFN) and transcription activator-like effector nucleases (TALEN) are one of the most promising tools for modifying genomes. These site-specific enzymes cause double- strand breaks that allow gene disruption or gene insertion, thereby facilitating genetic manipulation. The major problem associated with this approach is the labor-intensive procedures required to screen and confirm the cellular modification by nucleases. In this work, we produced a TALEN that targets the human CCR5 gene and developed a heteroduplex mobility assay for HEK 293T cells to select positive colonies for sequencing. This approach provides a useful tool for the quick detection and easy assessment of nuclease activity. |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: CCR5; Genome editing; Gene knockout; TALEN; HMA. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572014000100018 |
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