摘要
目的建立白藜芦醇合酶基因的毕赤酵母表达系统。方法通过PCR、限制性内切酶消化、连接、电激等方法,构建表达载体,转化毕赤酵母GS115。结果从葡萄基因组DNA中扩增得到了 1 200 bp目的基因,并克隆至pBS-T栽体;成功构建了表达载体pPIC3．5K／RS;目的基因成功整合进毕赤酵母GS115基因组中。测序及PCR鉴定证明,白藜芦醇合酶基因的毕赤酵母表达系统建立成功。结论所得方法无需高质量的RNA,从而降低了试验条件,达到了快速简便的目的。
Aim To construct the Pichia pastoris expression system of RS gene. Methods By PCR, enzyme di- gestion, ligation and electroporation, construct recombinant vector and transform Pichia pastoris GS115. Results The target fragment was amplified from the grape genome DNA and cloned into pBS-T Vector. ; The recombinant vector pPIC3.5K/RS was successfully constructed; The target gene was integrated into Pichia pastoris genome via homologous recombination . The results detected by sequencing and PCR show that the Pichia pastoris expression system of RS gene was successfully constructed. Conclusion The obtained method didnot reed high quality RNA, which lowered the experinment condition and made the process quick and simple.
出处
《西北大学学报（自然科学版）》
CAS
CSCD
北大核心
2006年第2期263-266,共4页
Journal of Northwest University（Natural Science Edition）
基金
陕西省自然科学基金资助项目(99JK122)
