Objective To establish a method of Campylobacter jejuni colonization in the intestine of C57BL/ 6 mice through antibiotic-induced microbiome depletion. Methods Thirty-six C57BL/ 6 female mice were divided into normal, control and experimental groups. After 2 days of cefoperazone sodium and sulbactam sodium intragastric administration (50 mg / mL), 200 μL of Campylobacter jejuni were administered intragastrically. 16S rDNA analysis was performed to detect intestinal contents on days 1 ~ 3 post modeling. Animal feces were collected, and the HipO gene of Campylobacter jejuni was detected by TaqMan qPCR on days 1 ~ 7 post modeling. Immunofluorescence was used to detect the colonization of Campylobacter jejuni in the intestinal tissues of mice at 1, 2, 3 and 7 days, and the pathological changes were observed by HE staining. Results In the experimental group, the Lactobacillus of the ileum and Bacteroides of the cecum and colon were inhibited. Additionally, the relative abundance of Enterococcus was increased, and a higher abundance of Campylobacter was detected on day 1. Higher Campylobacter jejuni copy numbers were detected in the feces of the experimental group by TaqMan qPCR on days 1~ 3, and immunofluorescence-labeled Campylobacter jejuni were clearly observed in the intestinal lumen. The intestinal mucosa was mainly intact and showed no obvious inflammatory cell infiltration. Conclusions Microbiome depletion induced by cefoperazone sodium and sulbactam sodium promoted the short- term colonization of Campylobacter jejuni in the intestine of mice.