Thioesterase catalyzes the hydrolysis of acyl-ACP and saturated fatty acyl chain. It plays a key role in the accumulation of medium chain fatty acids in vivo. In this study, to construct an engineering strain to produce MCFAs, the Arabidopsis acyl-ACP thioesterase gene AtFatA was amplified by PCR from cDNA of arabidopsis and double digested by EcoR I/Xba I, then linked to the plasmid digested with same enzymes to get the recombinant plasmid pPICZaA-AtFatA. We transformed the gene into Pichia pastoris GS115 by electroporation and screened positive colonies by YPD medium with Zeocin. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results showed that the recombinant enzyme had a molecular of 45 kDa band which was consistent with the predicted molecular mass and we constructed the expression system of gene AtFatA in fungus for the first time. Under shake-flask conditions, Gas Chromatograph-Mass Spectrometer-computer results indicated that recombinant strain produced 51% more extracellular free MCFAs than the wild and its yield reached 28.7% of all extracellular fatty acids. This figure is 10% higher than the control group. The result provides a new way to produce MCFAs.
Arabidopsis ; genetics ; Arabidopsis Proteins ; biosynthesis ; DNA, Complementary ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Pichia ; metabolism ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Thiolester Hydrolases ; biosynthesis ; Transformation, Genetic

Control at beyond-visual ranges is of great significance to animal-robots with wide range motion capability. For pigeon-robots, such control can be done by the way of onboard preprogram, but not constitute a closed-loop yet. This study designed a new control system for pigeon-robots, which integrated the function of trajectory monitoring to that of brain stimulation. It achieved the closed-loop control in turning or circling by estimating pigeons' flight state instantaneously and the corresponding logical regulation. The stimulation targets located at the formation reticularis medialis mesencephali (FRM) in the left and right brain, for the purposes of left- and right-turn control, respectively. The stimulus was characterized by the waveform mimicking the nerve cell membrane potential, and was activated intermittently. The wearable control unit weighted 11.8 g totally. The results showed a 90% success rate by the closed-loop control in pigeon-robots. It was convenient to obtain the wing shape during flight maneuver, by equipping a pigeon-robot with a vivo camera. It was also feasible to regulate the evolution of pigeon flocks by the pigeon-robots at different hierarchical level. All of these lay the groundwork for the application of pigeon-robots in scientific researches.
Animals ; Columbidae/physiology* ; Robotics/methods* ; Cerebral Cortex