The Antibody Preparation And The Study Of Its Effects On Dough Of HMW-GS 14 In Wheat

High-molecular-weight glutenins which constitute of many subunits is a kind of important storage protein and occupy 10% of whole protein in seed. Plenty of studies showed that constitution of high-molecular-weight glutenin subunit (HMW-GS)play a significant role on Triticeae processing quality. During the analysis of the contribute of different HMW-GS to processing quality find that 1Dx5+1Dy10 is quality subunits but 1Dx2+1Dy12 is poor subunits. Some quality bread Triticeae such as xiaoyan 6 (1Bx14+1By15 and 1Dx2+1Dy12)have no 1Dx5+1Dy10 but have 1Dx2+1Dy12 in wheat of our country. The analysis showed that 1Bx14+1By15 have the same function with 1Dx5+1Dy10, and research revealed 1Bx14+1By15 have significant effect to quality breeding. So studying the function of the subunit 1Bx14 which can have a profound effect on dough is a precondition to quality breeding. In this experiment, wheat xiaoyan6 was used as sample to find out a method that can recover and cumulate protein from SDS-PAGE effectively. Preparative electrophoresis(17cm*17cm*1.5mm) was applied to separate proteins in wheat. The gel was stained with KCl, then the part containing the aimed protein was cut from the gel and put in the dialytic sack for recovery. Results showed that subunits HMW14 and 15 can be separated more effectively by adding 4-vp during extraction process compared to normal method. SDS-PAGE showed that the purity of HMW14 subunit was single-point after 2-D electrophresis. In conclusion, this method supplies a new technique for effective separating of HMW-GS subunits in wheat. The development of reduction-oxidation methods has allowed new types of investigation of the effects of glutenin subunits on dough charaters. These methods can be used to research the native gluten polymer, in corporate monomers, and repolymerize the modified polymer. In this experiment, these methods were used to incorporate bulk high weight glutenin from four cultivar into base flours to detect its effects on dough quality. The protocol is as follows: the base flour is yuzhan 2000. The blends (flour and the protein fraction), 0.6mg of reducing agent DTT and distilled water were mixed for 30 sec. The partially reduced dough was treated with 7.2mg of oxidation (KIO3). Mixing was done in triplicate and parameters were recorded. The result showed that , after adding HMW14 subunits into the base flour the data of MT and PBR increased apprantly. In conclusion, HMW14 subunits have a positive effects on the dough quality.