|
|
|
| Construction of Glaesserella parasuis ptsG Mutant Strain and Analysis of Its Partial Biological Characteristics |
| YAN Xue-Feng1, XIAO Wu-Dian2, GU Cong-Wei2, HE Man-Li2, ZHAO Ming-De2, HE Lyu-Qin2,* |
1 School of Physical Education, Southwest Medical University, Luzhou 646000, China;
2 Laboratory Animal Center, Department of Science and Technology, Southwest Medical University, Luzhou 646000, China |
|
|
|
|
Abstract In the glucose-specific phosphotransferase (PTSGlc) system, glucose-specific enzyme ⅡBC (EⅡBCGlc) has a comprehensive effect on the metabolism of bacteria, but of which the function is still unclear in Glaesserella parasuis. In this study, EⅡBCGlc protein coding gene (ptsG) mutant and comlementary strain were constructed by natural transformation method. The growth status of wild strain SC1401 and ptsG mutant strains in the trypton soy broth (TSB)+glucose medium were compared in this study. Moreover, the bacterial morphology, biofilm formation, tolerance to oxygen pressure and virulence to mice (Mus musculus) of the SC1401 and ptsG mutant strain were measured. The results showed that the ptsG mutant strain could take in more reducing sugars for its own growth. Through transmission electron microscopy that compared with the wild strain, ptsG mutant strain had more high electron density areas inside the bacteria. In addition, the formation of biofilm and the tolerance to hydrogen peroxide were decreased in ptsG mutant strain. But there was no significant difference between SC1401 and ptsG mutant strains on mice. This study found that the ptsG gene played an important role in the formation of biofilm and oxidative stress tolerance, which will lay a foundation for the study of pathogenic mechanism in Glaesserella parasuis.
|
|
Received: 26 March 2021
|
|
|
|
Corresponding Authors:
*helvqin617@163.com
|
|
|
|
[1] 康磊, 李军星,王一成, 等. 2015. 副猪嗜血杆菌Wza基因缺失株的构建及生物学特性研究[J]. 农业生物技术学报,23(6):806-815.
(Kang L, Li J X, Wang Y C, et al.Construction and biological characteristic of Wza-deficient mutant of Haemophilus parasuis[J]. Journal of Agricultural Biotechnology, 23(6):806-815.)
[2] 林迪斯, 徐丽, 李飞雨, 等. 2018. 肺炎克雷伯菌果糖磷酸转移酶系统EⅡC编码基因frwC缺失株的构建及表型分析[J]. 西安交通大学学报(医学版), 39(6): 788-792.
(Ling D S, Xu L, Li F Y, et al.2018. Construction of mutant strain of Klebsiela pneumoniae fructose-specific phosphotransferase systemsubunit IIC encoded by frwC gene and analysis of its phenotypes[J]. Journal of Xi'an Jiaotong University (Medical Sciences), 39(6): 788-792.)
[3] 陆竞争, 任顺利, 诸葛斌, 等. 2017. 磷酸转移酶系统关键基因敲除对Klebsiella pneumoniae产1,3-丙二醇的影响[J]. 食品与发酵工业, 43(8): 26-30.
(Lu J Z, Ren S L, Zhu G B, et al.2017. The effects of knocking out key genes in phosphotransferase system on 1,3-propanediol production of Klebsiella pneumoniae[J]. Food and Fermentation Industries, 43(8): 26-30.)
[4] 万遂如. 2005. 副猪嗜血杆菌感染[J]. 养猪, 40-41.
(Wan S R.2005.Infection of Haemophilus parasuis[J]. Swine production , 40-41.
[5] 王欣然, 2012. 大肠杆菌BL21(DE3)和MG1655的代谢特征比较[D]. 硕士学位论文, 山东大学, 导师: 祁庆生. pp.1-91.
(Wang X R.2012. Comparision of the metabolic characteristics of Escherichia coli BL21 (DE3) and MG1655[D]. Thesis for M.S., Shandong University, Supervisor: Qi Q S, pp.1-91.)
[6] 吴永恒. 2017. 副猪嗜血杆菌病诊疗体会[J]. 中国畜禽种业, 122.
(Wu Y H.Experience in diagnosis and treatment of Haemophilus parasuis[J]. The Chinese Livestock and Poultry Breeding, 122.)
[7] 张旭, 2014. 葡萄糖专一性磷酸转移酶系统单基因缺失对大肠杆菌生理特性的影响[D]. 山东大学, 硕士学位论文, 山东大学, 导师: 祁庆生, pp. 1-83.
(Zhang X.2014. The effects of single gene deletion in the phosphoenolpyruvate-dependent glucose-specific phosphotransferase system on Escherichia coli physiological characters[D]. Thesis for M.S., Shandong University, Supervisor: Qi Q S.pp.1-83.)
[8] Ajdic D, Pham V T T.2017. Global transcriptional analysis of streptococcus mutans sugar transporters using microarrays[J]. Journal of Bacteriology, 189(14): 5049-5059.
[9] Deutscher J, Francke C, Postma P W.2006. How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria[J]. Microbiology & Molecular Biology Reviews, 70(4): 939-1031.
[10] Escalante A, Salinas Cervantes A, Gosset G, et al.2012. Current knowledge of the Escherichia coli phosphoenolpyruvate-carbohydrate phosphotransferase system: Peculiarities of regulation and impact on growth and product formation[J]. Applied Microbiology & Biotechnology, 94, 1483-1494.
[11] Gabor E, GÖhler A K, Kosfeld A, et al.2011. The phosphoenolpyruvate-dependent glucose-phosphotransferase system from Escherichia coli K-12 as the center of a network regulating carbohydrate flux in the cell[J]. European Journal of Cell Biology, 90(9): 711-720.
[12] Görke B, Stülke J, 2008. Carbon catabolite repression in bacteria: Many ways to make the most out of nutrients[J]. Nature Reviews Microbiology, 6(8): 613-624.
[13] Hasona A, Zuobi-Hasona K, Crowley P J, et al.2007. Membrane composition changes and physiological adaptation by Streptococcus mutans signal recognition particle pathway mutants[J]. J Bacteriol, 189(4): 1219-1230.
[14] He L, Dai K, Wen X, et al.2018. QseC mediates osmotic stress resistance and biofilm formation in Haemophilus parasuis[J]. Front Microbiol, 13(9): 212.
[15] Hengstenberg W, Egan J B, Morse M L.1967. Carbohydrate transport in Staphylococcus aureus. V. The accumulation of phosphorylated carbohydrate derivatives, and evidence for a new enzyme-splitting lactose phosphate[J]. Proceedings of the National Academy of Sciences of the USA, 58(1): 274-279.
[16] Hernández-Montalvo V, Martínez A, Hernández-Chavez G, et al.2003. Expression of galP and glk in a Escherichia coli PTS mutant restores glucose transport and increases glycolytic flux to fermentation products[J]. Biotechnology & Bioengineering, 83(6): 687-694.
[17] Jin H, Zhou R, Kang, M, et al.2006. Biofilm formation by field isolates and reference strains of Haemophilus parasuis[J]. Veterinary Microbiol, 118: 117-123.
[18] Kang Z, Geng Y, Xia Y Z, et al.2009. Engineering Escherichia coli for an efficient aerobic fermentation platform[J]. Journal of Biotechnology, 144(1): 58-63.
[19] Khan S R, Banerjeebhatnagar N, 2002. Loss of catabolite repression function of HPr, the phosphocarrier protein of the bacterial phosphotransferase system, affects expression of the cry4A toxin gene in Bacillus thuringiensis subsp. israelensis[J]. Journal of Bacteriology, 184(19): 5410-5417.
[20] Kotrba P, Inui M, Yukawa H, 2001. Bacterial phosphotransferase system (PTS) in carbohydrate uptake and control of carbon metabolism[J]. Journal of Bioscience & Bioengineering, 92(6): 502-517.
[21] Meins M, Jenö P, Müller D, et al.1993. Cysteine phosphorylation of the glucose transporter of Escherichia coli[J]. Journal of Biological Chemistry, 268(16): 11604-11609.
[22] Mijakovic I, Poncet S, Galinier A, et al.2002. Pyrophosphate-producing protein dephosphorylation by HPr kinase/phosphorylase: A relic of early life?[J]. Proceedings of the National Academy of Sciences of the USA, 99(21): 13442-13447.
[23] Mitchell W J, Saffen D.W, Roseman S, et al.1987. Sugar transport by the bacterial phosphotransferase system. In vivo regulation of lactose transport in Escherichia coli by IIIGlc, a protein of the phosphoenolpyruvate: Glycose phosphotransferase system[J]. Journal of Biological Chemistry, 262(33): 16254-16260.
[24] Nam T W, Jung H I, An Y J, et al.2008. Analyses of Mlc-IIBGlc interaction and a plausible molecular mechanism of Mlc inactivation by membrane sequestration[J]. Proceedings of the National Academy of Sciences of the USA, 105(10): 3751-3756.
[25] Oliveira S, Pijoan C.2004. Haemophilus parasuis: New trends on diagnosis, epidemiology and control[J]. Vet Microbiol, 99(1): 1-12.
[26] Phue J N, Shiloach J, 2004. Transcription levels of key metabolic genes are the cause for different glucose utilization pathways in E. coli B (BL21) and E. coli K (JM109)[J]. Journal of Biotechnology, 109: 21-30.
[27] Römling U, Sierralta W.D, Eriksson K Normark, et al.1998. Multicellular and aggregative behaviour of Salmonella typhimurium strains is controlled by mutations in the agfD promoter[J]. Molecular Microbiology 28(2): 249-264.
[28] Zhang B, Feng S, Xu, C, et al.2012. Serum resistance in Haemophilus parasuis SC096 strain requires outer membrane protein P2 expression[J]. FEMS Microbiology Letters, 326(2): 109-115.
[29] Zhang J, Xu C, Shen H, Li J, et al.2014. Biofilm formation in Haemophilus parasuis: Relationship with antibiotic resistance, serotype and genetic typing[J]. Research in Veterinary Science, 97(2): 171-175.
[30] Zogaj X, Nimtz M, Rohde M, et al.2001. The multicellular morphotypes of Salmonella typhimurium and Escherichia coli produce cellulose as the second component of the extracellular matrix[J]. Molecular Microbiology, 39(6):1452-1463. |
|
|
|
