关键词: avenin-like, 基因克隆, 载体构建, 融合蛋白, 原核表达

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.

Key words: Key words: avenin-like, gene cloning, vector construction, fusion protein, prokaryotic expression