• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >Targeted disruption of the NIT8 gene in Chlamydomonas reinhardtii.
【24h】

Targeted disruption of the NIT8 gene in Chlamydomonas reinhardtii.

机译：Reinhardtii衣藻中NIT8基因的靶向破坏。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

We have used homologous recombination to disrupt the nuclear gene NIT8 in Chlamydomonas reinhardtii. This is the first report of targeted gene disruption of an endogenous locus in C. reinhardtii and only the second for a photosynthetic eukaryote. NIT8 encodes a protein necessary for nitrate and nitrite assimilation by C. reinhardtii. A disruption vector was constructed by placing the CRY1-1 selectable marker gene, which confers emetine resistance, within the NIT8 coding region. nit8 mutants are unable to grow on nitrate as their sole nitrogen source (Nit-) and are resistant to killing by chlorate. One of 2,000 transformants obtained after selection on emetine-chlorate medium contained a homologous insertion of five copies of the disruption plasmid into the NIT8 gene, producing an emetine-resistant, chlorate-resistant Nit- phenotype. The mutant phenotype was rescued by the wild-type NIT8 gene upon transformation. Seven other mutations at the nit8 locus, presumably resulting from homologous recombination with the disruption plasmid, were identified but were shown to be accompanied by deletions of the surrounding genomic region.
机译：我们已经使用同源重组破坏莱茵衣藻中的核基因NIT8。这是针对雷氏梭菌内源基因座的靶向基因破坏的第一个报道，而光合作用的真核生物只有第二个报道。 NIT8编码莱茵衣藻对硝酸盐和亚硝酸盐吸收所必需的蛋白质。通过在NIT8编码区域内放置赋予曲美汀抗性的CRY1-1选择标记基因，构建了一个破坏载体。 nit8突变体无法作为唯一的氮源（Nit-）在硝酸盐上生长，并且耐氯酸盐杀死。在依米替丁-氯酸盐培养基上进行选择后获得的2,000个转化子之一，将5个拷贝的破坏质粒同源插入到NIT8基因中，产生了一个抗依米替丁，抗氯酸盐的Nit表型。转化后，野生型NIT8基因拯救了突变表型。确定了nit8基因座上的其他七个突变，可能是由于与破坏质粒的同源重组引起的，但显示有周围基因组区域的缺失。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号