Construction of a plasmid interspecific transfer system in Bacillus species with the counter-selectable marker mazF

Author:

Zhao XingYa12,Xu JianYong23,Tan Ming23,Yu ZhenXiao12,Yang ShiBin12,Zheng HongChen23,Song Hui123

Affiliation:

1. 0000 0004 1797 8419 grid.410726.6 University of Chinese Academy of Sciences 100049 Beijing China

2. 0000000119573309 grid.9227.e Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences 300308 Tianjin China

3. 0000000119573309 grid.9227.e Tianjin Key Laboratory for Industrial Biological, Systems and Bioprocessing Engineering, Tianjin Institute of Industrial Biotechnology Chinese Academy of Sciences 300308 Tianjin China

Abstract

Abstract Bacillus sp. strains as attractive hosts for the production of heterologous secretory proteins usually play important roles in bio-industry. However, low transformation efficiency of exogenous plasmids limited the application of Bacillus species. Here, a novel plasmid interspecific transfer system, with high transformation efficiency, high positive rate, and convenient manipulation, has been successfully constructed. A high electrotransformation efficiency strain Bacillus subtilis F-168 containing the counter-selectable marker mazF was used as the plasmid donor strain in this transfer method. A shuttled plasmid pBE980 and its recombinant plasmids pBE980::pulA and pBE980::HSPA were successfully transferred into the recipient Bacillus strains (Bacillus amyloliquefaciens 66, Bacillus licheniformis 124 and Bacillus megaterium 258) by this method. After co-culturing the donor cells (OD600nm = 1.3–1.7) and the recipient cells (OD600nm = 0.5–0.9) for 24 h in 22 °C, more than 1.0 × 105 positive transformants were obtained and a interspecific transformation efficiency of 1.0 × 10−3. It would provide a new approach for genetic manipulation in Bacillus strains and accelerate the research progress of the wild Bacillus strains in bio-industry.

Funder

the National Natural Science Fund of China

the Key Deployment Project in Chinese Academy of Sciences

the Tianjin Science & Technology Planning Project

Publisher

Oxford University Press (OUP)

Subject

Applied Microbiology and Biotechnology,Biotechnology,Bioengineering

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3