Construction of integrate plasmid pBlue-KanMX4-NTS2-5’3’-MSTN. (A) PCR amplification of ADH1 promoter (ADH1 p), Cyc3 termination signal (Cyc3 term) and the cDNA of myostatin (MSTN cDNA). (B) Overlapping PCR for myostatin expression cassette. (C) Restriction digestion of pBlue-KanMX4-NTS2-5’3’-MSTN with enzymes indicated in the figure. In panels (A) - (C), DNA fragments were separated in 1% agarose gel and visualized by ethidium bromide staining. DNA molecular weight marker was shown in lane 1. (D) Schematic illustration of integrate plasmid pBlue-KanMX4-NTS2-5’3’-MSTN.