https://ogma.newcastle.edu.au/vital/access/ /manager/Index en-au 5 https://ogma.newcastle.edu.au/vital/access/ /manager/Repository/uon:8253 Sat 24 Mar 2018 08:40:35 AEDT ]]> https://ogma.newcastle.edu.au/vital/access/ /manager/Repository/uon:34171 3 Ligand (FLT₃L) have many applications in cellular and molecular biology studies, and also as therapeutical agents. FLT₃L was expressed in Pichia pastoris through cloning from human leukemic K₅₆₂ cell line as well as using a codon-optimized synthetic construct. Codon adaptation index (CAI) was increased from 55 in the native sequence up to 95 after optimization. Significant changes occurred in the codons for Pro, Arg, Leu and Ser toward the favored codons in P. pastoris. Both forms of expressed FLT₃L (from the native and optimized sequences) were capable of stimulating proliferation of the FDC-P1 cells expressing human wild-type FLT₃ (FD-FLT₃-WT). Sequence optimization resulted in 55-fold increase in the yield of active FLT₃L (290 µg/mL of product in the crude supernatant). Amino acid residues 27-162 are sufficient for the biological function of human FLT₃L. Pichia pastoris is a highly efficient and cost-effective system for expression of endotoxin-free FLT₃L; however, codon optimization is necessary for its optimal expression.]]> Mon 18 Feb 2019 13:59:00 AEDT ]]>