关键词: 植物乳杆菌, 亚油酸异构酶, 克隆, 生物信息学

Abstract: According to the nucleotide sequence of linoleate isomerase gene published in GenBank, a pair of specific primers of linoleate isomerase was designed. The target gene was amplified by PCR and cloned into T vector, and the recombinant plasmid was then transformed into Trans1-T1 competent cells. Based on the colony PCR identification authentication and sequence analysis, the length of linoleate isomerase gene was 1720 bp. The target gene had 99% homology with linoleate isomerase gene of Lactobacillus plantarum AS1.555 (DQ227322) and 89% homology with L. plantarum kimchi (DQ010331). The sequence had been registered in Genbank as the number of HM569265. Bioinformatic analysis exhibited the garget gene had a segment with low composition complexity. By aligning genome sequence of Lactobacillus (CP001617), specific primers of gene sequence with 805 bp located at the upstream non-coding region of linoleate isomerase gene were designed, and then the sequence was amplified by PCR and cloned into T vector, and the recombinant plasmid was then transformed into Trans1-T1 competent cells. Colony PCR identification, preliminary bioinformatic analysis and sequence analysis revealed 12 motifs and 6 transcription factor binding sites of linoleate isomerase gene.

Key words: Lactobacillus plantarum, linoleate isomerase, cloning, bioinformatic analysis