A chimeric vacuolar Na+/H+ antiporter gene evolved by DNA family shuffling confers increased salt tolerance in yeast
文献类型：SCI/SSCI论文
| 作者 | Wu G. X.; Wang, G.; Ji, J.; Li, Y.; Gao, H. L.; Wu, J.; Guan, W. Z. |
| 发表日期 | 2015 |
| 关键词 | DNA family shuffling Na+/H+ antiporter Salt tolerance Yeast hydrophilic c-terminus arabidopsis-thaliana salicornia-europaea directed evolution expression plants sos1 thermostability recombination transporters |
| 英文摘要 | The vacuolar Na+/H+ antiporter plays an important role in maintaining ionic homeostasis and the osmotic balance of the cell with the environment by sequestering excessive cytoplasmic Na+ into the vacuole. However, the relatively low Na+/H+ exchange efficiency of the identified Na+/H+ antiporter could limit its application in the molecular breeding of salt tolerant crops. In this study, DNA family shuffling was used to create chimeric Na+/H+ antiporters with improved transport activity. Two homologous Na+/H+ antiporters from halophytes Salicornia europaea (SeNHX1) and Suaeda salsa (SsNHX1) were shuffled to generate a diverse gene library. Using a high-throughput screening system of yeast complementation, a novel chimeric protein SseNHX1 carrying 12 crossover positions and 2 point mutations at amino acid level was selected. Expression of SseNHX1 in yeast mutant exhibited approximately 46% and 22% higher salt tolerance ability in yeast growth test than that of SsNHX1 and SeNHX1, respectively. Measurements of the ion contents demonstrated that SseNHX1 protein in yeast cells accumulated more Na+ and slightly more K+ than the parental proteins did. Furthermore, this chimera also conferred increased tolerance to LiC1 and a similar tolerance to hygromycin B compared with the parental proteins in yeast. (C) 2015 Elsevier B.V. All rights reserved. |
| 出处 | Journal of Biotechnology |
| 卷 | 203 |
| 页 | 1-8 |
| 收录类别 | SCI |
| 语种 | 英语 |
| ISSN号 | 0168-1656 |
| 源URL | [http://ir.igsnrr.ac.cn/handle/311030/38944]  |
| 专题 | 地理科学与资源研究所_历年回溯文献 |
| 推荐引用方式 GB/T 7714 | Wu G. X.,Wang, G.,Ji, J.,et al. A chimeric vacuolar Na+/H+ antiporter gene evolved by DNA family shuffling confers increased salt tolerance in yeast. 2015. |
入库方式： OAI收割
来源：地理科学与资源研究所
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