Journal: International Journal of Molecular Sciences
Article Title: Engineering of Fatty Acid Synthases (FASs) to Boost the Production of Medium-Chain Fatty Acids (MCFAs) in Mucor circinelloides
Figure Lengend Snippet: ( A , B ) Expression of TE-01 , TE-02 , TE-03 , and TE-04 genes. ( A ) Structure of plasmids pMAT1552-pyrF-TE-01, pMAT1552-pyrF-TE-02, pMAT1552-pyrF-TE-03, pMAT1552-pyrF-TE-04 for TE-01 , TE-02 , TE-03 , TE-04 genes over-expressing in M. circinelloides M65 are demonstrated. Green boxes indicate the coding region of thioesterase genes. ( B ) Polymerase chain reaction (PCR) amplification of genome of control strain (A) and thioesterase gene (TE) over-expressing strains i.e., M65-TE-01, M65-TE-02, M65-TE-03, and M65-TE-04 was demonstrated to as B–E, respectively with the primers ( Supplementary materials: Table S2 ). M, Gene Ruler DNA Ladder Mix. Sizes in kb of the relevant maker fragments are indicated.
Article Snippet: The aforesaid primers exhibited homologous sequences (i.e., 28 bp) to the Xho I restriction sites in plasmid pMAT1552-pyrF, and the PCR fragment was finally cloned into plasmid pMAT1552-pyrF using the restriction endonuclease Xho I to construct the recombinant plasmids i.e., pMAT1552-pyrF-TE-01, pMAT1552-pyrF-TE-02, pMAT1552-pyrF-TE-03, pMAT1552-pyrF-TE-04 (One-Step Cloning Kit from Takara Bio USA, Inc., CA, USA).
Techniques: Expressing, Polymerase Chain Reaction, Amplification