马铃薯反义SSU原核表达载体的构建及其对大肠杆菌糖原合成的影响.pdfVIP

安徽农业科学，Jo~nal of Anhui Agri．Sci．2007，35(36)：11762～11763 责任编辑 孙红忠 责任校对 李洪 马铃薯反义SSU原核表达载体的构建及其对大肠杆菌糖原合成的影响 万巧凤 ，姚新灵孙，丁海麦 (1．宁夏医学院高职学院，宁夏银川750021；2．宁夏大学生命科学学院，宁夏银川750021；3．包头医学院基础部，内蒙古包头014010) 摘要 [目的]为了鉴定马铃著反义ssu的功能。[方法]利用分子生物学技术，构建了马铃薯反义ssu eDNA原核表达栽体(OFtaS)，通过 0．1 mol／L llrlG诱导pF~-aS在大肠杆菌BI21(DE3)中表达，测定糖原的相对含量。[结果]经连接、转化和酶切鉴定筛选出重组表达裁体 oFtaS，而pFtaS转化BI21(OE3)获得重组菌株BI21一aS。经WIG诱导的BI21一aS糖原含量OD值为0．409，显著低于对照菌株BI21(DD) 和未经IlrrG诱导的BI21一aS，而对照菌株BI21(OE3)和未经llrlG诱导的BI21一aS的糖原含量OD值差异不显著。这表明反义SSU在BL21 内表达能够抑制glgC编码的AGPase的形成，从而减少其糖原合成量。[结论]该研究为构建马铃薯反义SSU植物转化栽体奠定了基础。 关键词 AGPase；原核表达载体；糖原；植物转化栽体 中图分类号 Q5O3 文献标识码 A 文章编号 0517—6611(2OO7)36—11762—02 Censtnletion ofPotato Antlsense ssU Prokaryotlc Expression Vector and Its Effect Oil file Glycogen Synthesis of E．coil Qiao．feng et al(flier Vocational School ofl~ingxia Medical College，Yinchuan，l~ingxia750021) Abstract【ObjectiveJ 1k research aimed to diSellS8 tlle function of potato antisenese SSU、【Metl~]By moleodar biological techniques，珊矗 expression vector(oFtaS)for potota antisenese SSU eDNAwas constructed．AndpE-aSwas induced to express in E．eoli BI21(OE3)by0．1 mol／LIFIV, to deterrffmethe relative con~nt ofglycogen．【ResultJ Recombinant expression vectorpE-aSwas screened outthrou~connection，transfonmtion ar,d既一 zyme digestion identificati~．And recombinant$1~dill BI2l—aS was obtained from transforming BI2l(oE3)by pE-aS．1he 01)value of yc0靴 content in B~2l—aStIllou 研Ginducti~was 0．409，si can由 Iowerthanthatin contmI$1~dillBL21(DE3)aridB【2l—aSwithout I’Ginduetio~a． ．dettae OD values of幽 啪content between control strain BI2l(oE3)and BL21一aS had no significant difference．This indicated that t} expression ofarllJse． nese SSU in BL21 could inhibit tlle fonmtion of AGPase coded by gtgc to reduce tlle synthetic quantity of glycogen．『Conclusion]Ittis research laid a fomldationfor c0fls衄lc planttransfonmtion vectorfor potato antisenese SSU． Key 咖 s A