Establishment of a markerless gene deletion system in Chromohalobacter salexigens DSM 3043@

文献类型： 外文期刊

第一作者： Shao, Ya-Hui

作者： Shao, Ya-Hui;Guo, Li-Zhong;Yu, Hao;Lu, Wei-Dong;Zhao, Bai-Suo

作者机构：

关键词： Chromohalobacter salexigens;Allelic exchange;Homologous recombination;Markerless deletion;sacB

期刊名称：EXTREMOPHILES （ 影响因子：2.395； 五年影响因子：2.668 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Chromohalobacter salexigens DSM 3043 can grow over a wide range of salinity, which makes it as an excellent model organism for understanding the mechanism of prokaryotic osmoregulation. Functional analysis of C. salexigens genes is an essential way to reveal their roles in cellular osmoregulation. However, the lack of an effective markerless gene deletion system has prevented construction of multiple gene deletion mutants for the members in the genus. Here, we report the development of a markerless gene deletion system in C. salexigens using allelic exchange method. In this system, the in vitro mutant allele of target gene was inserted into a pK18mobsacB-based integrative vector pMDC21, which contained a chloramphenicol resistance cassette as the positive selection marker and a sacB gene from Bacillus subtilis as the counterselectable marker. To validate this system, two single-gene deletion mutants and a double-gene deletion mutant were constructed. In addition, our results showed that growth of the merodiploids and sucrose screening at 25 degrees C were more effective to decrease the occurrence of spontaneous sucrose resistance colonies than at higher temperature (30 or 37 degrees C), and growth of the merodiploids in mineral salt medium instead of the complex medium was critical to increase the recovery rate of deletion mutants.

分类号： Q93

  • 相关文献

[1]New genetic mechanism, origin and population dynamic of bovine ephemeral fever virus. He, Cheng-Qiang,Liu, Ya-Xin,Ding, Nai-Zheng,Wang, Hong-Mei,Hou, Pei-Li,He, Hong-Bin.

[2]An enhanced vector-free allele exchange (VFAE) mutagenesis protocol for genome editing in a wide range of bacterial species. Gomaa, Ahmed E.,Zhang, Chen,Yang, Zhimin,Shang, Liguo,Jiang, Shijie,Deng, Zhiping,Zhan, Yuhua,Lu, Wei,Lin, Min,Yan, Yongliang. 2017

[3]Variation between Turnip mosaic virus isolates in Zhejiang Province, China and evidence for recombination. Chen, JP,Chen, JP,Adams, MJ. 2002

[4]The Klebsiella pneumoniae nitrogenase Fe protein gene (nifH) functionally substitutes for the chlL gene in Chlamydomonas reinhardtii. Cheng, Q,Day, A,Dowson-Day, M,Shen, GF,Dixon, R. 2005

[5]Construction of a Bacillus thuringiensis engineered strain with high toxicity and broad pesticidal spectrum against coleopteran insects. Liu, Jingjing,Yan, Guixin,Shu, Changlong,Zhao, Can,Song, Fuping,Zhou, Lin,Ma, Junlan,Zhang, Jie,Liu, Chunqin,Huang, Dafang.

[6]Construction of recombinant Marek's disease virus (MDV) lacking the meq oncogene and co-expressing AIV-H9N2 HA and NA genes under control of exogenous promoters. Zhang, Zhenjie,Chen, Wenqing,Ma, Chengtai,Zhao, Peng,Duan, Luntao,Zhang, Fushou,Su, Hongqin,Li, Sifei,Cui, He,Cui, Zhizhong,Sun, Aijun,Li, Yanpeng.

[7]Crossover rate between homologous chromosomes and interference are regulated by the addition of specific unpaired chromosomes in Brassica. Suay, Loreto,Zhang, Deshuang,Eber, Frederique,Jouy, Helene,Lode, Maryse,Huteau, Virginie,Coriton, Olivier,Szadkowski, Emmanuel,Paillard, Sophie,Chevre, Anne-Marie,Zhang, Deshuang,Leflon, Martine,Martin, Olivier C.,Falque, Matthieu,Jenczewski, Eric.

[8]High-Frequency Targeted Mutagenesis in Pseudomonas stutzeri Using a Vector-Free Allele-Exchange Protocol. Gomaa, Ahmed E.,Deng, Zhiping,Yang, Zhimin,Shang, Liguo,Zhan, Yuhua,Lu, Wei,Lin, Min,Yan, Yongliang.

[9]Impact of rli87 gene deletion on response of Listeria monocytogenes to environmental stress. Xie Kun,Meng Qingling,Jun Qiao,Peng Yelong,Liu Tianli,Chen Cheng,Ma Yu,Hu Zhengxiang,Chen Chuangfu,Cai Xuepeng.

[10]A simplified system for generating recombinant E3-deleted canine adenovirus-2. Yu, Zuo,Jiang, Qian,Liu, Jiasen,Guo, Dongchun,Quan, Chuansong,Li, Botao,Qu, Liandong. 2015

[11]A ku70 null mutant improves gene targeting frequency in the fungal pathogen Verticillium dahliae. Qi, Xiliang,Qi, Juncang,Qi, Xiliang,Su, Xiaofeng,Guo, Huiming,Cheng, Hongmei.

[12]A novel approach for the construction of plant amiRNA expression vectors. Yan, Hong,Deng, Xuan,Ma, Lixin,Yan, Hong,Zhao, Bin,Cao, Yinglong,Huang, Junyan.

[13]Evolution and homologous recombination of the hemagglutinin-esterase gene sequences from porcine torovirus. Cong, Yingying,Wang, Xin,Suo, Siqingaowa,Ren, Yudong,Ren, Xiaofeng,Zarlenga, Dante S.,Richt, Juergen A.,Wang, Yang,Wang, Jingfei,Wang, Jingfei.

[14]The potential mechanism of Bursal-derived BPP-II on the antibody production and avian pre-B cell. Feng, Xiuli,Cao, Ruibing,Zhou, Bin,Liu, Ke,Liu, Xiaodong,Zhang, Yuanpeng,Gu, Jinyan,Chen, Puyan,Liu, Qingtao,Miao, Denian. 2013

[15]Efficient gene replacements in ku70 disruption strain of Aspergillus chevalieri var. intermedius. Huang, Qingqing,Huang, Qingqing,Cao, Yang,Liu, Zuoyi,Tan, Yumei,Liu, Yongxiang. 2017

[16]Highly Efficient CRISPR/Cas9-Mediated Homologous Recombination Promotes the Rapid Generation of Bacterial Artificial Chromosomes of Pseudorabies Virus. Guo, Jin-Chao,Tang, Yan-Dong,Zhao, Kuan,Wang, Tong-Yun,Liu, Ji-Ting,Gao, Jia-Cong,Chang, Xiao-Bo,Cui, Hong-Yu,Tian, Zhi-Jun,Can, Xue-Hui,An, Tong-Qing. 2016

[17]Altering fibroin heavy chain gene of silkworm Bombyx mori by homologous recombination. Zhao, Y,Chen, X,Peng, WP,Dong, L,Huang, JT,Lu, CD. 2001

[18]Co-Transfer of TALENs Construct Targeted for Chloroplast Genome and Chloroplast Transformation Vector Into Rice Using Particle Bombardment. Li, Ding,Tang, Ning,Fang, Zhen,Xia, Yumei,Cao, Mengliang,Li, Ding,Tang, Ning.

[19]Phylogenetic analysis of VP1 gene sequences of waterfowl parvoviruses from the Mainland of China revealed genetic diversity and recombination. Wang, Shao,Cheng, Xiao-xia,Chen, Shao-ying,Lin, Feng-qiang,Chen, Shi-long,Zhu, Xiao-li,Wang, Jin-xiang,Huang, Mei-qing,Zheng, Min,Wang, Shao,Cheng, Xiao-xia,Chen, Shao-ying,Lin, Feng-qiang,Chen, Shi-long,Zhu, Xiao-li,Wang, Jin-xiang,Huang, Mei-qing,Zheng, Min.

[20]Enhanced gene replacement frequency in KU70 disruption strain of Stagonospora nodorum. Feng, Jie,Hwang, Sheau-Fang,Li, Weijiao,Gossen, Bruce D.,Strelkov, Stephen E..

作者其他论文 更多>>
  • Comparison of plastisphere microbiomes during the degradation of conventional and biodegradable mulching films

    作者：Yu, Hao;Zhang, Mingyang;Zhang, Mingyang;Xiao, Juanjuan;Men, Jianan;Deng, Ye;Jin, Decai;Liu, Huiying;Cernava, Tomislav;Cernava, Tomislav

    关键词：Biodegradable mulching film; Plastisphere; Bacterial community; Soil microbiome; Assembly processes

  • The impact of nozzle spacing on the vortex evolution and dilution characteristics of pulsed jets in crossflow

    作者：Wang, Chuan;Wang, Chuan;Yu, Hao;Zhang, Li;Wang, Chuan;Wang, Chuan;Li, Hao;Zhang, Li

    关键词：Nozzle spacing; Pulse; Triple; Counter-rotating vortex pair; Diffusion

  • OsPAD1, encoding a non-specific lipid transfer protein, is required for rice pollen aperture formation

    作者：Wang, Qiming;Tian, Yunlu;Chen, Keyi;Wang, Chaolong;Yu, Xiaowen;Bai, Wenting;Zheng, Hai;You, Shimin;Hu, Yang;Lei, Dekun;Jian, Anqi;Lu, Jiayu;Yu, Hao;Jiang, Ling;Zhao, Zhigang;Wan, Jianmin;Zhu, Shanshan;Xiong, Yehui;Zhang, Xin;Ren, Yulong;Lei, Cailin;Cheng, Zhijun;Lin, Qibing;Wan, Jianmin

    关键词：Sterile; Fibrillar-granular layer; Operculum; Lipid binding; Oriza sativa

  • OrchidBase 6.0: increasing the number of Cymbidium (Orchidaceae) genomes and new bioinformatic tools for orchid genome analysis

    作者：Chen, You-Yi;Sun, Ye;Li, Chung-, I;Lin, Shao-Ting;Yang, Chien-Wen;Tsai, Wen-Chieh;Lin, Shao-Ting;Yang, Chien-Wen;Tsai, Wen-Chieh;Zheng, Hao-Chen;Zhang, Zhe-Bin;Lee, Bing-Ru;Hsieh, Chun-Lin;Wu, Wei-Sheng;Hsiao, Yu-Yun;Tsai, Wen-Chieh;Chow, Chi-Nga;Chang, Wen-Chi;Tsai, Wen-Chieh;Chen, Heming;Yang, Feng-Xi;Zhu, Gen-Fa;Zheng, Qinyao;Zhou, Cheng-Yuan;Zhao, Zhuang;Ai, Ye;Wang, Lin-Ying;Peng, Dong-Hui;Lan, Siren;Liu, Zhong-Jian;Zheng, Qinyao;Zhou, Cheng-Yuan;Zhao, Zhuang;Ai, Ye;Wang, Lin-Ying;Peng, Dong-Hui;Lan, Siren;Liu, Zhong-Jian;Chen, Deqiang;He, Xin;Huang, Ming-Zhong;Yu, Hao;Tsai, Wen-Chieh;Tsai, Wen-Chieh

    关键词：Orchid; OrchidBase; Cymbidium; Whole-genome sequencing; Genome; Transcriptome

  • EasyMetagenome: A user-friendly and flexible pipeline for shotgun metagenomic analysis in microbiome research

    作者：Bai, Defeng;Xun, Jiani;Ma, Chuang;Luo, Hao;Yang, Haifei;Hou, Huiyu;Lv, Hujie;Wan, Xiulin;Wang, Yao;Yousuf, Salsabeel;Zeng, Meiyin;Zhang, Tianyuan;Gao, Yunyun;Liu, Yong-Xin;Chen, Tong;Ma, Chuang;Yang, Haifei;Cao, Chen;Cao, Xiaofeng;Cui, Jianzhou;Deng, Yuan-Ping;Deng, Zhaochao;Yu, Hao;Zhang, Chunfang;Dong, Wenxin;Dong, Wenxue;Du, Juan;Fang, Qunkai;Fang, Wei;Fang, Yue;Luan, Yaning;Fu, Fangtian;Fu, Min;Fu, Yi-Tian;Gao, He;Ge, Jingping;Guo, Yuhao;Gong, Qinglong;Lou, Wenbo;Gu, Lunda;Yang, Li;Guo, Peng;Hai, Tang;Liu, Hao;He, Jieqiang;He, Zi-Yang;Huang, Can;Ji, Shuai;Jiang, ChangHai;Jiang, Gui-Lai;Jiang, Lingjuan;Jin, Ling N.;Li, Changchao;Kan, Yuhe;Kang, Da;Kou, Jin;Lam, Ka-Lung;Li, Chong;Li, Fuyi;Li, Liwei;Li, Miao;Li, Xin;Li, Ye;Li, Zheng-Tao;Zhu, Chengshuai;Liang, Jing;Mo, Jiayuan;Lin, Yongxin;Liu, Changzhen;Liu, Danni;Zhang, Jing;Chen, Shifu;Liu, Fengqin;Liu, Jia;Liu, Tianrui;Liu, Tingting;Wang, Xinlong;Liu, Xinyuan;Luo, Yuanyuan;Liu, Yaqun;Liu, Bangyan;Liu, Minghao;Lv, Hujie;Ma, Tengfei;Mai, Zongjiong;Niu, Dongze;Pan, Zhuo;Qi, Heyuan;Shi, Zhanyao;Song, Chunjiao;Sun, Fuxiang;Sun, Yan;Tian, Sihui;Wang, Guoliang;Wang, Hongyang;Wang, Hongyu;Wang, Huanhuan;Wang, Jing;Wang, Jun;Wang, Kang;Wang, Leli;Yao, Xiaofang;Wang, Shao-kun;Xiao, Zufei;Xing, Huichun;Xu, Yifan;Yang, Song;Yan, Shu-yan;Zhang, Yi-Bo;Yang, Yuanming;Lei, Yu;Yuan, Zhengrong;Zhang, Chunge;Zhang, Huimin;Zhang, Na;Zhang, Yupeng;Zhang, Zheng;Zhou, Mingda;Zhou, Yuanping;Zhu, Zhihao;Zhu, Lin;Zhu, Yue;Zou, Hongqin;Zuo, Anna;Dong, Wenxuan;Wen, Tao;Chen, Shifu;Chen, Shifu;Li, Guoliang

    关键词：metagenome; microbiome; microbiota; pipeline; visualization

  • Multiomic Analysis Provided Insights into the Responses of Carbon Sources by Wood-Rotting Fungi Daldinia carpinicola

    作者：Yang, Peng;Han, Jiandong;Wan, Luzhang;Yang, Peng;Han, Jiandong;Wan, Luzhang;Ma, Xingchi;Zhang, Yu;Sun, Yanan;Ma, Meng;Cheng, Fansheng;Zhang, Yu;Yu, Hao;Cheng, Fansheng

    关键词：wood-rotting fungi; CAZyme; carbon source; sucrose; sawdust; secondary metabolites

  • Proteomics and physiologic analysis reveal different response strategies to cadmium stress in Lentinula edodes

    作者：Wu, Yansha;Zhai, Dandan;Jiang, Ning;Li, Qiaozhen;Zhang, Meiyan;Song, Chunyan;Dong, Haoran;Shang, Xiaodong;Yu, Hailong;Zhai, Dandan;Yu, Hailong;Yu, Hao

    关键词：Cadmium; Lentinula edodes; Glutathione; Proline; Proteomics

微信小程序