摘要
[目的]为了鉴定马铃薯反义SSU的功能。[方法]利用分子生物学技术,构建了马铃薯反义SSUcDNA原核表达载体(pE-aS),通过0.1mol/LIPTG诱导pE-aS在大肠杆菌BL21(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载体pE-aS,而pE-aS转化BL21(DE3)获得重组菌株BL21-aS。经IPTG诱导的BL21-aS糖原含量OD值为0.409,显著低于对照菌株BL21(DE3)和未经IPTG诱导的BL21-aS,而对照菌株BL21(DE3)和未经IPTG诱导的BL21-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植物转化载体奠定了基础。
[Objective] The research aimed to discuss the function of potato antisenese SSU.[Method] By molecular biological techniques,prokaryotic expression vector（pE-aS）for potota antisenese SSU cDNA was constructed.And pE-aS was induced to express in E.coli BL21（DE3） by 0.1 mol/L IPTG to determine the relative content of glycogen.[Result] Recombinant expression vector pE-aS was screened out through connection,transformation and enzyme digestion identification.And recombinant strain BL21-aS was obtained from transforming BL21（DE3） by pE-aS.The OD value of glycogen content in BL21-aS through IPTG induction was 0.409,significantly lower than that in control strain BL21（DE3） and BL21-aS without IPTG induction.While the OD values of glycogen content between control strain BL21（DE3） and BL21-aS had no significant difference.This indicated that the expression of antisenese SSU in BL21 could inhibit the formation of AGPase coded by glgC to reduce the synthetic quantity of glycogen.[Conclusion] This research laid a foundat
ion for constructing plant transformation vector for potato antisenese SSU.
出处
《安徽农业科学》
CAS
北大核心
2007年第36期11762-11763,共2页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(30160010)
教育部科学研究重点研究课题
宁夏自然科学研究基金项目(C132)
关键词
AGPASE
原核表达载体
糖原
植物转化载体
AGPase
Prokaryotic expression vector
Glycogen
Plant transformation vector
