...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Intein-mediated assembly of a functional beta -glucuronidase in transgenic plants.
【24h】

Intein-mediated assembly of a functional beta -glucuronidase in transgenic plants.

机译：内蛋白介导的转基因植物中功能性β-葡萄糖醛酸苷酶的组装。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The DnaE intein in Synechocystis sp. strain PCC6803 is the first and only naturally split intein that has been identified so far. It is capable of catalyzing a protein trans-splicing mechanism to assemble a mature protein from two separate precursors. Therefore, it is a powerful tool for protein modification and engineering. Inteins have not been identified, nor have intein-mediated protein splicing reactions been demonstrated, in plant cells. In this paper, we describe the use of the Ssp DnaE split intein in transgenic plants for reconstitution of a protein trans-splicing reaction. We have synthesized artificial genes that encode for N-terminal half (Int-n) and C-terminal half (Int-c) fragments of Ssp DnaE split intein and divided beta-glucuronidase (GUS) gene to encode GUS-n and GUS-c parts of the enzyme as reporter. The in-frame fusions of GUSnIntn and IntcGUSc were constructed and transfected into Arabidopsis. We have observed in vivo reassembly of functional beta-glucuronidase when both GUSnIntn and IntcGUSc constructs were introduced into the same Arabidopsis genome either by cotransformation or through genetic crossing, hereby signifying an intein-mediated protein trans-splicing mechanism reconstituted in plant cells.
机译：Synechocystis sp。中的DnaE内含子。菌株PCC6803是迄今已鉴定出的第一个也是唯一的天然分裂内含子。它能够催化蛋白质反式剪接机制，以从两个单独的前体组装成熟蛋白质。因此，它是蛋白质修饰和工程化的强大工具。在植物细胞中，尚未鉴定出内含肽，也未证实内含肽介导的蛋白质剪接反应。在本文中，我们描述了Ssp DnaE分裂内含子在转基因植物中重组蛋白反式拼接反应的用途。我们已经合成了人工基因，该基因编码Ssp DnaE拆分的Intein的N端一半（Int-n）和C端一半（Int-c）片段，并分割了β-葡萄糖醛酸苷酶（GUS）基因来编码GUS-n和GUS- c部分酶为报告基因。构建了GUSnIntn和IntcGUSc的框内融合物，并将其转染到拟南芥中。当通过共转化或通过遗传杂交将GUSnIntn和IntcGUSc构建体引入相同的拟南芥基因组时，我们已经观察到功能性β-葡糖醛酸糖苷酶的体内重组，由此表明在植物细胞中重构的内含肽介导的蛋白质反式剪切机制。

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