Optimization of plant des-pGlu1-Brazzein gene according to yeasty biased codons and its expression in Pichia pastoris.
- Author:
Chunli LI
1
;
Lu HAN
;
Zhenyu ZHENG
;
Weidong ZHAO
Author Information
1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China. hnclli@163.com
- Publication Type:Journal Article
- MeSH:
Codon;
Pichia;
genetics;
metabolism;
Plant Proteins;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification;
Sweetening Agents
- From:
Chinese Journal of Biotechnology
2011;27(8):1158-1163
- CountryChina
- Language:Chinese
-
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.
