P158: Name tagging of lactic acid bacteria by random integration of DNA barcode

DNA barcodes is a short sequence of intrinsic DNA that can be easily generated and characterized for all species on the planet. In this way, DNA barcoding is usually used for large surveys aiming at unknown species detection and identification of species with medical, ecological and agronomical significance. Here, we intentionally incorporated a synthesized DNA barcode in the genome of lactic acid bacteria for marking bio-information. The DNA barcode was comprised of sequences having specific PCR primer, alphabetical information sequence of bacterial strain, chloramphenicol-resistant gene (CAT), and mosaic end (ME) sites for recognition of transposase. Microbial strains, owners, inventers and the number of patent depository were encoded in the base sequence. The constructed DNA barcode was randomly integrated into chromosomal DNA of Leuconostoc citreum HJ-P4 using a viral transposase after electroporation. The chloramphenicol resistant mutants were isolated and successful integration of barcode sequence was confirmed by PCR amplification using designed primers. Mutants integrated DNA barcode had same growth pattern with Leuconostoc citreum HJ-P4, but little slow. Also, DNA barcode stability of mutants was maintained at 80% until 1000 generations. For application of DNA barcode in food, HJ-P4 integrated DNA barcode were inoculated as the starter in kimchi. DNA barcode was monitored by PCR-DGGE method during kimchi fermentation. This barcoding system can be used to recognize, detect and trace dispersal of patented organisms in biotechnology, either to certify the source organism or secure intellectual property rights for bioresources.