...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Codon optimization and expression of irisin in Pichia pastoris GS115
【24h】

Codon optimization and expression of irisin in Pichia pastoris GS115

机译：毕赤酵母GS115中虹膜素的密码子优化和表达

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

Irisin is a novel hormone which is related to many metabolic diseases. In order to illuminate the function and therapeutic effect of irisin, gaining active irisin is necessary. In this work, a codon-optimized irisin gene was designed according to Pichia pastoris synonymous codon usage bias and cloned into the pPIC9K expression vector. Sequencing result indicating that the sequence of irisin was consistent with the modified irisin and the irisin was in frame with a-factor secretion signal ATG. The plasmid pPIC9K-irisin was transformed into GS115 P. pastoris cells through electroporation. The positive transformants were screened on MD medium and analyzed by PCR. Five recombinant GS115/pPIC9K-irisin strains were obtained, but only one strain expressed irisin successfully. SDS-PAGE and Western blot were used to assess the expression level and purity of irisin. The irisin was also simply purified and the effect of pH value, methanol concentration and induction time on the production of irisin was investigated. The results showed that the best conditions of irisin expression were as follows: pH 6.0, 2.0% methanol and induction for 96h. This work laid the basis for further investigation into the therapeutic and pharmacological effects of irisin, as well as development of irisin-based therapy. (C) 2015 Elsevier B.V. All rights reserved.
机译：鸢尾素是一种新型激素，与许多代谢性疾病有关。为了阐明虹膜素的功能和治疗效果，必须获得活性虹膜素。在这项工作中，根据巴斯德毕赤酵母同义密码子使用偏倚设计了密码子优化的虹膜素基因，并将其克隆到pPIC9K表达载体中。测序结果表明，鸢尾素的序列与修饰的鸢尾素一致，且该鸢尾素与α因子分泌信号ATG在读框内。通过电穿孔将质粒pPIC9K-irisin转化到GS115巴斯德毕赤酵母细胞中。在MD培养基上筛选阳性转化体，并通过PCR进行分析。获得了5株重组GS115 / pPIC9K-irisin菌株，但只有1株成功表达了虹膜素。 SDS-PAGE和Western印迹用于评估虹膜蛋白的表达水平和纯度。虹膜素还可以简单纯化，并研究pH值，甲醇浓度和诱导时间对虹膜素生产的影响。结果表明，虹膜素表达的最佳条件如下：pH 6.0，甲醇2.0％，诱导96h。这项工作为进一步研究虹膜素的治疗和药理作用以及开发基于虹膜素的疗法奠定了基础。 （C）2015 Elsevier B.V.保留所有权利。

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