Amplification of 16SrDNA and mecA genes
According to the instruction of DNA extraction kit, the genomic DNA of the isolated bacterial strain was extracted. The primers (Forward:5’-AGAGTTTGATCCTGGCTCAG-3’, Reverse:5’-GGTTACCTT GTTACGACTT -3’) and the conditions (95 C for 1 min; 30 cycles of 95 C for 30s, 55 C for 30 s and 72 C for 90s followed by 1 cycle of 72 C for 10 min) were used for amplifying 16SrDNA gene with the size of 1465 bp; the primers (forward:5’-GTAGAAATGACTGAACGTCCGATAA-3’; Reverse :5’-CCAATTCCACATTGTTTCGGT CTAA-3’) and the conditions (95 C for 1 min; 30 cycles of 95 C for 30s, 55 C for 30 s and 72 C for 90s, followed by 1 cycle of 72 C for 10 min) were used for amplifying mecA gene with the size of 310 bp. The PCR products were observed by 1% agarose gel electrophoresis, and then were sequenced by Beijing Liuhe Huada Genome Technology Co., Ltd.
The 16SrDNA sequence of the isolated bacterial strain was compared with that of the published other strains in GenBank of NCBI, and its homology and phylogenetic tree were analyzed using DNA Star MEGA7 software.