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OALib Journal期刊
ISSN: 2333-9721
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Optimization of plant des-pGlu1-Brazzein gene according to yeasty biased codons and its expression in Pichia pastoris
植物甜蛋白des-pGlu1-Brazzein的密码子优化及其在毕氏酵母中的表达

Keywords: des-pGlu1-Brazzein,Pichia pastoris,expression,biological activity
甜蛋白,毕氏酵母,表达,活性检测

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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.

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