Z Fan, H B Liu, Y C Chen, X H Cui, Z F Li, T T Fu, J Yuan
{"title":"[肺炎克雷伯氏菌中 wza 基因缺失对胶囊形成和噬菌体敏感性的影响]。","authors":"Z Fan, H B Liu, Y C Chen, X H Cui, Z F Li, T T Fu, J Yuan","doi":"10.3760/cma.j.cn112150-20240226-00148","DOIUrl":null,"url":null,"abstract":"<p><p><b>Objective:</b> To investigate the effects of <i>wza</i> gene deletion in <i>Klebsiella pneumoniae</i> on capsule formation ability and bacteriophage sensitivity. <b>Methods:</b> The <i>wza</i> deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ<i>wza</i> were detected by measuring the optical density OD<sub>600</sub>. In order to analyze the effect of gene <i>wza</i> on bacterial capsule formation, wild-type strain W14 and Δ<i>wza</i> mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ<i>wza</i> mutant strain. The <i>t</i> test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ<i>wza</i> mutant strain. <b>Results:</b> The PCR results revealed that the Δ<i>wza</i> mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ<i>wza</i> on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ<i>wza</i> mutant strains in LB broth. The transmission electron microscope results showed that <i>wza</i> gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ<i>wza</i> mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups (<i>t</i>=27.233, <i>P<</i>0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene <i>wza</i> was deleted. <b>Conclusion:</b> Deletion of the <i>wza</i> gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.</p>","PeriodicalId":24033,"journal":{"name":"中华预防医学杂志","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-07-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[The effect of <i>wza</i> gene deletion in <i>Klebsiella pneumoniae</i> on capsule formation and bacteriophage sensitivity].\",\"authors\":\"Z Fan, H B Liu, Y C Chen, X H Cui, Z F Li, T T Fu, J Yuan\",\"doi\":\"10.3760/cma.j.cn112150-20240226-00148\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Objective:</b> To investigate the effects of <i>wza</i> gene deletion in <i>Klebsiella pneumoniae</i> on capsule formation ability and bacteriophage sensitivity. <b>Methods:</b> The <i>wza</i> deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ<i>wza</i> were detected by measuring the optical density OD<sub>600</sub>. In order to analyze the effect of gene <i>wza</i> on bacterial capsule formation, wild-type strain W14 and Δ<i>wza</i> mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ<i>wza</i> mutant strain. The <i>t</i> test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ<i>wza</i> mutant strain. <b>Results:</b> The PCR results revealed that the Δ<i>wza</i> mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ<i>wza</i> on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ<i>wza</i> mutant strains in LB broth. The transmission electron microscope results showed that <i>wza</i> gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ<i>wza</i> mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups (<i>t</i>=27.233, <i>P<</i>0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene <i>wza</i> was deleted. <b>Conclusion:</b> Deletion of the <i>wza</i> gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.</p>\",\"PeriodicalId\":24033,\"journal\":{\"name\":\"中华预防医学杂志\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-07-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华预防医学杂志\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3760/cma.j.cn112150-20240226-00148\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华预防医学杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3760/cma.j.cn112150-20240226-00148","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
[The effect of wza gene deletion in Klebsiella pneumoniae on capsule formation and bacteriophage sensitivity].
Objective: To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods: The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δwza were detected by measuring the optical density OD600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δwza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δwza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δwza mutant strain. Results: The PCR results revealed that the Δwza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δwza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δwza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δwza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups (t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion: Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.
期刊介绍:
Chinese Journal of Preventive Medicine (CJPM), the successor to Chinese Health Journal , was initiated on October 1, 1953. In 1960, it was amalgamated with the Chinese Medical Journal and the Journal of Medical History and Health Care , and thereafter, was renamed as People’s Care . On November 25, 1978, the publication was denominated as Chinese Journal of Preventive Medicine . The contents of CJPM deal with a wide range of disciplines and technologies including epidemiology, environmental health, nutrition and food hygiene, occupational health, hygiene for children and adolescents, radiological health, toxicology, biostatistics, social medicine, pathogenic and epidemiological research in malignant tumor, surveillance and immunization.