Abstract: 【Objective】The present study aimed at cloning, prokaryotic expression and functional analysis of avenin-like genes from common wheat.【Method】The avenin-like genes were cloned by gene specific primers, which designed according to conserved domains of published genes, followed by sequencing and bioinformatic anayzing. Hereafter, JN542444 was cloned into the expression vector, and transformed into the host bacteria Escherichia coli Rosetta-gami B(DE3). After induced by IPTG, the fusion protein was purified by His-Trap affinity chromatography, and then integrated into the control flour to determine the quality effect utilization of 10 g Farinograph. 【Result】 Twenty-two novel avenin-like genes (GenBank accession No. were from JN542443 to JN542464) with 855 bp were isolated by gene-specific primers from 12 wheat cultivars, and all of them encoded 284 amino acid residues. The number and the positions of Cys residues were strictly conservative in 21 genes, which possibly could form 7 intra- and 4 inter-disulfide bonds, except JN542452 with 17 Cys residues. Three pseudogenes, including JN542448, JN542455 and JN542456, were resulted from a single base substitution. An unique type of the avenin-like gene, which was identical to 7 different cultivars in this study and event the other cultivars in GenBank, was named “avenin-like I”. Phylogenetic and evolutionary analysis revealed that avenin-like subunits were closely related to LMW-GS and least homology with ω-gliadin in wheat storage proteins. SDS-PAGE resuslts showed that the gene was successfully expressed in E. coli system. The farinograph data of purified protein revealed that avenin-like I has a significant positive effect on wheat dough quality.【Conclusion】Although lack of polymorphism in the tested cultivars, the Avenin-like I subunit could improve wheat flour processing quality.

Key words: Triticum aestivum L., avenin-like protein, prokaryotic expression, quality effect