数字农科院2.0

Highly Effective Markerless Genetic Manipulation of Streptococcus suis Using a Mutated PheS-Based Counterselectable Marker

文献类型： 外文期刊

作者： Guangjuan Gao;Dong Wei;Gang Li;Ping Chen;Liujun Wu;Siguo Liu;Yueling Zhang

作者机构：

关键词： counterselectable marker;markerless genetic manipulation;mutated PheS;Streptococcus suis;strong promoter

期刊名称： Frontiers in Microbiology

ISSN： 1664-302X

年卷期： 2022 年 13 卷

页码：

摘要： Streptococcus suis is an important zoonotic pathogen, however, an efficient markerless genetic manipulation system is still lacking for further physiological and pathological studies on this bacterium. Several techniques have been developed for markerless genetic manipulation of S. suis utilizing either a temperature-sensitive vector or a counterselectable markers (CSMs), however, at present, the efficiency of these techniques is not very satisfactory. In this study, we developed a strategy for markerless genetic manipulation of S. suis employing a CSM based on a conditionally lethal mutant allele of pheS, which encodes the α-subunit of phenylalanyl-tRNA synthetase (PheS). This mutant pheS, mPheS, was constructed by introducing site-directed mutations for a T261S/A315G double-substitution and a number of silent mutations to decrease its similarity with the endogenous wild type pheS gene (wtPheS). Additionally, five potentially strong promoters from S. suis were screened for their ability to drive high-level expression of mPheS, thus endowing the carrier strain with sufficient sensitivity to the phenylalanine analog p-chloro-phenylalanine (p-Cl-phe). Insertion of these P-mPheS cassettes into a vector or into the chromosomal locus via a linked erythromycin resistance gene revealed that mPheS allele driven by promoters P0530 and P1503 renders S. suis sensitive to as low as 0.01% (or 0.5 mM) of p-Cl-phe. This offers two potential CSMs for S. suis with p-Cl-phe as a counterselective agent. P1503-mPheS was revealed to be 100% efficient for counter-selection in S. suis by application in a precise gene deletion. Using P1503-mPheS as a CSM, a two-step insertion and excision strategy for markerless genetic manipulation of S. suis were developed, supplying a powerful tool for markerless genetic manipulation of S. suis.

分类号：

  • 相关文献

[1]Gene Expressions in Major Organs between Enshi Black and Landrace Pigs after Streptococcus suis Infection. Gaur, Uma,Wu, Hua-Yu,Feng, Zheng,Yang, Xue-Hai,Guo, Rui,Wu, Jun-Jing,Mei, Shu-Qi,Liu, Gui-Sheng,Gaur, Uma,Wu, Hua-Yu,Feng, Zheng,Yang, Xue-Hai,Guo, Rui,Wu, Jun-Jing,Mei, Shu-Qi,Liu, Gui-Sheng,Li, Kui,Jin, Mei-Lin.

[2]Genome-wide identification of allele-specific expression in response to Streptococcus suis 2 infection in two differentially susceptible pig breeds. Wu, Huayu,Gaur, Uma,Peng, Xianwen,Li, Lianghua,Sun, Hua,Song, Zhongxu,Dong, Binke,Li, Mingbo,Mei, Shuqi,Liu, Guisheng,Wu, Huayu,Gaur, Uma,Peng, Xianwen,Li, Lianghua,Sun, Hua,Dong, Binke,Mei, Shuqi,Liu, Guisheng,Mekchay, Supamit,Wimmers, Klaus,Ponsuksili, Siriluck,Li, Kui.

[3]Erythromycin Resistance of Streptococcus suis Isolates In Vivo. Jiang Cheng-gang,Cai Xue-hui,Qiu Hua-ji,Li Yan-hua,Tong Heng-min,Liu Di-qiu,Lei Lian-cheng,Sun Chang-jiang. 2009

[4]Characterization of Streptococcus suis isolates from the diseased pigs in China between 2003 and 2007. Wei, Zigong,Li, Ran,Zhang, Anding,He, Hongkui,Hua, Yafeng,Xia, Jing,Chen, Huanchun,Jin, Meilin,Wei, Zigong,Li, Ran,Zhang, Anding,He, Hongkui,Hua, Yafeng,Xia, Jing,Chen, Huanchun,Jin, Meilin,Cai, Xuehui.

[5]GAP-initiated constitutive expression of a novel plectasin-derived peptide MP1106 by Pichia pastoris and its activity against Streptococcus suis. Jiao, Jian,Feng, Xingjun,Jiao, Jian,Mao, Ruoyu,Wang, Xiumin,Zhang, Yong,Teng, Da,Wang, Jianhua,Jiao, Jian,Mao, Ruoyu,Wang, Xiumin,Zhang, Yong,Teng, Da,Wang, Jianhua. 2015

[6]NeuA O-acetylesterase activity is specific for CMP-activated O-acetyl sialic acid in Streptococcus suis serotype 2. Song, Lili,Zhou, Hui,Jin, Cheng,Cai, Xuehui,Li, Chunyang,Liang, Jingnan. 2011

[7]Genetic diversity and virulence of novel sequence types of Streptococcus suis from diseased and healthy pigs in China. Wang, Shujie,Gao, Mingming,An, Tongqing,Liu, Yonggang,Jin, Jiamin,Wang, Gang,Jiang, Chenggang,Tu, Yabin,Hu, Shouping,Cai, Xuehui,Li, Jinsong,Wang, Jie,Zhou, Dongsheng. 2015

[8]Pdh Modulate Virulence Through Reducing S.tress Tolerance And Biofilm F ormation Of Streptococcus Suis Serotype 2. Wang, Y, Wang, YX, Liu, BB, Wang, SH, Li, JP, Gong, SL, Sun, LY, Yi, L. 2019

[9]Interaction between Porcine Alveolar Macrophage-Tang Cells and Streptococcus suis Strains of Different Virulence: Phagocytosis and Apoptosis. Li S.,Wang C.,Tang Y.-D.,Qin L.,Chen T.,Wang S.,Bai Y.,Cai X.,Wang S.. 2023

[10]Specific quantitative detection of Streptococcus suis and Actinobacillus pleuropneumoniae in co-infection and mixed biofilms. Yi, Li,Jin, Manyu,Gao, Mengxia,Wang, Haikun,Fan, Qingying,Grenier, Daniel,Sun, Liyun,Wang, Shaohui,Wang, Yang. 2022

[11]Establishment and Application of an Indirect ELISA for the Detection of Antibodies to Porcine Streptococcus suis Based on a Recombinant GMD Protein. Nihua Dong,Zhaofei Wang,Qing Sun,Xiaojun Chen,Hailong Zhang,Jiayang Zheng,Xinya Zhang,Yafeng Qiu,Zongjie Li,Beibei Li,Ke Liu,Donghua Shao,Jianchao Wei,Jianhe Sun,Zhiyong Ma. 2023

[12]New Characterization of Multi-Drug Resistance of Streptococcus suis and Biofilm Formation from Swine in Heilongjiang Province of China. Dong C.-L.,Che R.-X.,Wu T.,Qu Q.-W.,Chen M.,Zheng S.-D.,Cai X.-H.,Wang G.,Li Y.-H.. 2023

[13]A Unique Combination Of Glycoside Hydrolases In Streptococcus Suis Specifically And Sequentially Acts On Host-Derived Alpha Gal-Epitope Glycans. Chen, P, Liu, R, Huang, MM, Zhu, JL, Wei, D, Castellino, FJ, Dang, GH, Xie, F, Li, G, Cui, ZY, Liu, SG, Zhang, YL. 2020

[14]Viral Coinfection Replaces Effects Of S.uilysin On Streptococcus Suis A dherence To And Invasion Of Respiratory Epithelial Cells Grown Under Air-Liquid Interface Conditions. Meng, FD, Tong, J, Votsch, D, Peng, JY, Cai, XH, Willenborg, M, Herrler, G, Wu, NH, Valentin-Weigand, P. 2019

[15]Antimicrobial Susceptibility, Serotypes, And Genetic D.iversity Of Streptococcus Suis I n Diseased And Healthy Pigs In Southern China. Peng, L, Xu, CB, Yang, XF, Liu, BT. 2019

[16]In Vitro/Vivo Mechanism Of Action O.f Mp1102 With Low/Nonresistance A gainst Streptococcus Suis Type 2 Strain Cvcc 3928. Zhao, F, Yang, N, Wang, XM, Mao, RY, Hao, Y, Li, ZZ, Wang, X, Teng, D, Fan, H, Wang, JH. 2019

[17]GrpE and ComD contribute to the adherence, biofilm formation, and pathogenicity of Streptococcus suis. Fei Yu,Chunliu Dong,Yuefeng Zhang,Ruixiang Che,Chunmei Xie,Yanyan Liu,Zhiyun Zhang,Lu Li,Xueying Chen,Xuehui Cai,Gang Wang,Yanhua Li. 2023

[18]Subunit Vaccine Targeting Phosphate ABC Transporter ATP-Binding Protein, PstB, Provides Cross-Protection against Streptococcus suis Serotype 2, 7, and 9 in Mice. Yan Z.,Yao X.,Pan R.,Zhang J.,Ma X.,Dong N.,Wei J.,Liu K.,Qiu Y.,Sealey K.,Nichols H.,Jarvis M.A.,Upton M.,Li X.,Ma Z.,Liu J.,Li B.. 2023

[19]Streptococcus suis contributes to inguinal lymph node lesions in piglets after highly pathogenic porcine reproductive and respiratory syndrome virus infection. Wang, Shujie,Xu, Min,Yang, Kongbin,Zhang, Ying,Li, Siqi,Tang, Yan-Dong,Wang, Jinliang,Leng, Chaoliang,An, Tongqing,Cai, Xuehui. 2023

[20]Bovine Herpesvirus-4 Based Vaccine Provides Protective Immunity against Streptococcus suis Disease in a Rabbit Model. Nihua Dong,Hester Nichols,Qing Sun,Xiaojun Chen,Jiayang Zheng,Zhixin Guan,Hailong Zhang,Andrew Davison,Yvonne Wezel,Zongjie Li,Beibei Li,Ke Liu,Donghua Shao,Yafeng Qiu,Jianhe Sun,Xiangdong Li,Mathew Upton,Zhiyong Ma,Michael A. Jarvis,Jianchao Wei. 2023

作者其他论文 更多>>