FOOD SCIENCE ›› 2011, Vol. 32 ›› Issue (17): 297-302.doi: 10.7506/spkx1002-6630-201117062

• Bioengineering • Previous Articles     Next Articles

Cloning and Analysis of Linoleate Isomerase Gene from Lactobacillus plantarum P8 and Its Upstream Non-coding Region

ZHANG Xin-yan1,ZHAO Guo-fen1,*,BAO Qiu-hua2,ZHANG He-ping2   

  1. (1. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China; 2. College of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China)
  • Published:2011-08-30

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

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