华北农学报2013，Vol.28Issue(1)：101-105,5.
小麦ATG8的原核表达及其抗血清制备
Cloning and Prokaryotic Expression of ATG8 (a Molecular Marker of Autophagy)Gene from Wheat and Preparation of Antiserum
摘要
Abstract
Total RNA was isolated from wheat variety L10 and reverse transcription by using the anchored primers in this paper to further understand the biological function of the autophagy gene ATG8 from wheat. And then the ATG8 were amplified by PCR and then cloned into the PMD19-T vector,one of which was subcloned into the expression vector. The recombinant plasmid was identified by sequencing and digestion of restriction enzymes. The re-combinant expression vector was constructed and transformed into E. coli strain Roseta ( DE3 ) subsequently, then through IPTG-induction in host bacteria E.coli Rosetta-gami B(DE3)and detected by SDS-PAGE. The rabbit anti-ATG8 antibody was prepared and was detected by Western Blotting analysis. The ATG8 were obtained partly and successfully expressed in the prokaryotic expression system. The experiment offered foundation to the studying of autophagy and function of ATG8 gene in wheat.关键词
ATG8/免疫印迹/抗血清/融合蛋白/原核表达Key words
ATG8/Western Blotting/Anti-serum/Fusion protein/Prokaryotic expression分类
农业科技引用本文复制引用
吴洪波,刘刚,张路路,王冬梅..小麦ATG8的原核表达及其抗血清制备[J].华北农学报,2013,28(1):101-105,5.基金项目
国家自然科学基金(30671244 （）
31171472) （）
河北省应用基础研究计划重点基础研究项目(08965505D) （08965505D）
河北省自然科学基金(C2007000515 （）
C2010000787) （）
