title: [Optimization of plant des-pGlu1-Brazzein gene according to yeasty biased codons and its expression in Pichia pastoris].
abstract: According to the amino acid sequence of des-pGlu1-Brazzein, 4 pairs of oligonucleotide with cosmic site were synthesized by using yeasty biased codons. After linkage and PCR, the 179 bp code area of des-pGlu1-Brazzein was obtained and inserted into pPIC9K, which resulted in the recombinant expression vector pPIC9K-Bra. By digestion with Sal I, the lined pPIC9K-Bra was transformed into Pichia pastoris GS115 by electric shock. The results of expression indicted that the secreted target protein accounted for 51.6% of total protein in the supernatant and showed biological activity after purification.
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China. hnclli@163.com
authors.forename: Chunli
authors.initials: C
authors.lastname: Li
authors.forename: Lu
authors.initials: L
authors.lastname: Han
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Zheng
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhao
mesh_terms: D003062:Codon; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013549:Sweetening Agents
chemical_list: D003062:Codon; D010940:Plant Proteins; D011994:Recombinant Proteins; D013549:Sweetening Agents; C090521:brazzein protein, Pentadiplandra brazzeana