一株粗糙型牛种布鲁氏菌诱导株的构建及鉴定

doi:10.16431/j.cnki.1671-7236.2020.11.004

Abstract

Abstract: The purpose of the experiment was to study a new development method of brucellosis vaccine,and to successfully induce a rough Brucella abortus attenuated strain named RB71 by op inducer.Deletions of RB71-related genes were detected,The LPS integrity,growth characteristics,genetic stability,and viability in murine macrophage RAW264.7 cells were compared with those in smooth strains.The results showed that the mutant was successfully induced in the test,and the size of the missing fragment was 15 070 bp.The heat agglutination test was positive,which could be stained by crystal violet,and the acridine yellow agglutination test was positive.The extraction of lipopolysaccharide for silver staining showed that the O chain was deleted,and the induced strain RB71 lipopolysaccharide was incomplete.No gene mutation was detected by PCR after 30 consecutive passages.Under the same culture conditions in vitro,the growth rate of the induced strain RB71 was significantly lower than that of the parent strain A19.When infected with RAW264.7 macrophages in mice for 72 h,the intracellular survival rate was significantly reduced than that of the parent strain (P<0.01).In summary,this experiment successfully obtained an induced strain of Brucella RB71 with good genetic stability through the op inducer mutagenesis technique the survival ability of the induced strain in RAW264.7 cells was significantly weakened,which laid the technical foundation for the development of a new attenuated Brucella rough vaccine.

Key words: induce; rough; Brucella; vaccines

CLC Number:

S852.61

SUN Haojie, REN Xiaoxia, QIN Yuming, ZHU Liangquan, JIANG Hui, SUN Shijing, DING Jiabo, XIN Lingxiang, WANG Nan, LI Xiaoning, LI Qiaoling, MAO Kairong, CAI Yanan, XU Lei. Construction and Identification of an Induced Strain of Rough Brucella abortus[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3445-3453.

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Construction and Identification of an Induced Strain of Rough Brucella abortus

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