摘要
将本实验室已成功构建的植酸酶基因表达载体pPIC9K-phyA酶切线性化后,通过原生质体法整合到巴氏毕赤酵母(Pichia pastoris)GS115菌株细胞中,利用测定酶活、观察转化子在MM和MD平板上的生长情况以及PCR菌落鉴定,筛选出表型为甲醇利用缓慢型的重组子(His+Muts)。在相同培养条件下比较甲醇利用缓慢型的重组子和Mut+重组子在表达植酸酶方面的异同。实验结果表明,诱导表达48h后,Mut+和MutS重组子表达的产物在SDS-PAGE胶上都出现了清晰的目的带。酶学性质测定表明,酶学性质无显著差异。
The expression vector pPIC9K including phytase gene was linearized and then transformed into Pichia pastoris GS115 by means of producing plasmatosome. The Mut^s recombinant was screened by phenotype and PCK and the expression of phytase was compared with Mut^＋ recombinant in Pichia pastoris under the same condition. SDS-PAGE results indicated that there was a clear target band in Mut^s and Mut^＋ recombinant Culture supernatant after 48 hours culture respectively. Enzyme properties of tests showed that there was no significant difference between them.
出处
《生物技术通报》
CAS
CSCD
2006年第3期58-62,共5页
Biotechnology Bulletin
