摘要
设计合成3条引物,以载体pcDNA3.1-hAchE为模板,扩增出两端带有EcoRⅠ的约1.8 kb 去除信号肽但保留18 bp分子内伴侣序列的hAchE结构基因,及去除信号肽序列与分子内伴侣序列的hAchE成熟肽基因。将两个基因插入到酵母分泌型表达载体pPIC9和pHIL-S1,获得正向插入的4个分泌型酵母表达载体。为转化酵母,筛选分泌型高效表达hAchE菌株奠定了基础。
Two hAchE DNA fragments with EcoRⅠsites in distal end were amplified by PCR using 3 primers and the template in pcDNA3.1-hAchE. One of the fragments, the structural gene, comprised 18 bp molecular partners, the other was a mature peptide gene with deletions of the signal peptide and the 18 bp molecular partners. The DNA fragments were inserted into pPIC9 vector and pHIL-S1 vector, respectively, 4secretive expression vectors of pichia pastoris with human acetylcholinesterase (hAchE) gene were got. The study laid foundation for the screening of high expression of recombinant hAchE in yeast.
出处
《黑龙江八一农垦大学学报》
2004年第1期65-68,共4页
journal of heilongjiang bayi agricultural university
基金
全军医药卫生"十五"重点科研基金项目(01Z023)。
