Molecular Identify Of Wheat Avenin-like B Gene From Transgenic Plants

Wheat Avenin-like protein is a new type of seed storage proteins. The sequence ofAvenin-like protein was predicted by the sequence of avenin-like gene, rich in cysteineresidues. Avenin-like protein is speculated to affect processing quality of wheat flour.Therefore, studying on the relationship between Avenin-like protein and wheatprocessing quality will help to reveal the mechanism of wheat processing quality.In this study, the objective of this research was transgenic wheat plants which willbe detected at the DNA and protein molecular level. Southern blot was used to detectavenin-like b gene copy numbers of transgenic plants and progeny plants. And Westernblot technique was used to detect the expression of Avenin-like b protein in transgenicplants seeds. We obtained exogenous avenin-like b gene copies and overexpression intransgenic wheat plants. The results of present research provide materials for furtherstudy on the relationship between avenin-like b gene and processing quality. In thisresearch, we got the main results as follows:（1）The avenin-like b gene was amplified and then was cloned into vector pET32ato construct prokaryotic expression vector pET32a-avel. Avenin-like b gene wassuccessfully expressed in Escherichia coli strain BL21. Avenin-like b protein waspurified by affinity chromatography techniques.The purified protein was used toimmunize white rabbit and polyclonal antibodies were raised.（2）The copy numbers of avenin-like b gene were detected by Southern blottechnique. The results of Southern blot demonstrated that the avenin-like b gene wassuccessfully integrated into the genome of transgenic plants.（3）These antibodies were used to detect the expression of Avenin-like b proteinsin endosperms in transgenic plants by Western blot. The results showed that theexpression of Avenin-like b protein in transgenic plants increased significantly comparedto the wild-type plants. In this study, the transgenic plants and progeny plants were confirmed at the DNAand protein molecular level by Southern blot and Western blot. We obtained avenin-like bgene copies and overexpression in transgenic wheat plants, which provide materials forfurther study on the relationship between avenin-like b gene and wheat processingquality.