摘要
应用PCR技术从具有分泌蛋白能力强且没有胞外蛋白酶活性的短短小芽孢杆菌 5 0中分离出细胞壁蛋白基因的多启动子和信号肽编码序列 ,利用它与质粒pUB110和pKF3一起构建成穿梭分泌表达载体pBKE5 0。将α 淀粉酶基因引入该载体转化短短小芽孢杆菌 5 0后 ,发现α 淀粉酶可以活性形式分泌表达。此工作为下一步建立短短小芽孢杆菌高效分泌表达系统奠定了基础。
The 5′ region of the cell wall protein(CWP) gene containing multiple tandem promoters and the signal peptide-coding sequence was isolated by PCR from Br. brevis 50, and used to construct the shuttle vector pBKE50,which included the replication origin of pUB110 and the erythromycin\|resistance gene of pGK12.The α\|amylase gene of Bacillus subtilis 168 was ligated to pBKE50, producing plasmid pBKE50/α-amy.After the resulting plasmid was introduced into Br.brevis 50,soluble and biologically active α-amylase was secreted directly into the culture medium. The expression level of α-amylase in the recombinant Br.brevis 50 was twice higher than that of the donor strain.
出处
《生物工程学报》
CAS
CSCD
北大核心
2002年第4期438-441,共4页
Chinese Journal of Biotechnology
基金
军事医学科学院创新课题启动基金资助课题 (No .0 0 10 0 18)~~
