【摘要】 【目的】克隆小麦品种陕253中的新型avenin-like(类燕麦储藏蛋白),并构建该基因的原核表达载体,在大肠杆菌中诱导表达融合蛋白。【方法】利用PCR方法从小麦品种陕253中克隆新型avenin-like,将克隆的avenin-like亚克隆至表达载体,经酶切及测序鉴定后,转化宿主菌Escherichia coli Rosetta-gami B(DE3),经IPTG诱导表达融合蛋白,并利用Western blot进行检测。【结果】从小麦品种陕253克隆获得3个新avenin-like,GenBank登录号分别为GQ903577、GQ903578和GQ903579。序列分析表明,其中GQ903579为假基因,利用所构建的大肠杆菌表达载体,经IPTG诱导,实现了对登录号为GQ903578的基因在原核系统的特异性表达。【结论】新型avenin-like的克隆及原核表达的成功,为小麦品质改良提供了研究基础。更多还原

【Abstract】 Objective Molecular cloning and sequence characterization analysis of avenin-like from wheat cultivar Shaan 253, which were then constructed their prokaryotic expression vector and expressed in Escherichia coli Rosetta-gami B (DE3). Method The novel avenin-like were amplified from wheat variety Shaan 253 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 fusion protein was finally expressed by IPTG-induction in host bacteria-E. coli Rosetta-gami B (DE3) and detected by Western blot. Result Three novel avenin-like were obtained, and submitted to GenBank, and their accession No. are GQ903577, GQ903578, and GQ903579, respectively. Sequence analysis revealed that GQ903579 was a pseudogene. The gene GQ903578 was successfully expressed in the prokaryotic expression system. Conclusion It could provide a foundation for quality improvement of wheat.更多还原