Fig. 1.

Evaluation of transformation through PCR. a The sbtA and bicA gene constructs, having promoter and antibiotic selection marker were incorporated between NSI and NSII by double homologous recombination. Arrows indicate the primers used for PCR amplification of the region between NSI and NSII sites. WT, genomic DNA from the wild-type cells was used as the template; M, molecular weight marker; A, genomic DNA from strain A (sbtA-overexpressing strain) was used as the template; pA, plasmid pA was used as the template; B, genomic DNA from strain B (bicA-overexpressing strain) was used as the template; pB, plasmid pB was used as the template; NTC, no-template control, only buffer used as the template. b Measurement of relative levels of sbtA and bicA mRNA using qRT-PCR. N = 3. c Measurement of the expression of SbtA and BicA proteins using Western blotting using an anti-His6 antibody. L, Protein marker; NC, negative control, proteins extracted from the wild-type cells; A, proteins extracted from strain A; B, proteins extracted from strain B