Abstract: Objective: Lactobacillus acidophilus produces bacteriocin, bacteriocin gene was cloned and sequence was analyzed.Methods: The isolated strain was identified by 16 S rRNA.Primers were designed based on the reports plnD gene, the bacteriocin related genes of Lactobacillus acidophilus zrx02 was amplified by PCR and sequenced, then the bioinformatics were determined using online software ( Pro Param tool, TMHMM) . Results: The strain was identified as Lactobacillus acidophilus and named Lactobacillus acidophilus zrx02.The gene essential for plnD was present in Lactobacillus acidophilus zrx02, which was naturally clustered from Lactobacillus acidophilus in Gen Bank, the gene sequence of the plnD gene exhibited high similarity up to 100% to that of Lactobacillus plantarum strain ( WP 016526896.1) plnD gene.The results of sequencing showed that the full length of the plnD gene was 355 bp, which coded for 92 amino acid residues, with molecular weight of 10.54 ku, theoretical PI of 9.26, plnD encoded protein was stable protein. In addition, there was no obvious transmembrane structure present in the encoded product.The secondary structure of protein consists of α-helix, random coil and extended strand. The protein tertiary structure exhibited high similarity to ComE from Streptococcus pneumonia. Conclusion: The cloning and analysis of plnD gene in Lactobacillus acidophilus zrx02 are helpful to further explore of bacteriocin production.