GmDof4 transformation vector and detection of GmDof4 and npt II in the transformants. (A) A schematic map of the pGmDof4 plasmid. (B) PCR and RT-PCR detection of transgenic strains. Top panel: A 903 bp full length GmDof4 cDNA sequence was amplified in transgenic strains by PCR; Middle panel: A 773 bp fragment of GmDof4 cDNA was found in transgenic strains by RT-PCR; Bottom panel: A 623 bp fragment of nptII was found in transgenic strains by RT-PCR. (C) Southern blot detection of transgenic GmDof4 Chlorella ellipsoidea. The genomic DNA was digested with Hind III and Xba I or with Hind III and Nco I and then hybridized with a 545 bp fragment of the partial GmDof4 gene, which was labelled with α-32P dCTP by Random Primer DNA Labeling Kit ver. 2.0 (Takara Biotechnology Co.,Ltd., Da Lian, China). Dof4-1, Dof4-3, and Dof4-5: different transgenic GmDof4 strains. CK: pCK transgenic strains; PC: positive control; M: DNA molecular weight marker; WT: wild-type Chlorella ellipsoidea.