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Function Analysis of BL0033 and BL0034 in Fructose ABC Transporter System of Bifidobacterium Longum

Author: GuoYanHong
Tutor: LiaoXiangRu
School: Jiangnan University
Course: Biochemistry and Molecular Biology
Keywords: Fructose transport system BL0033 BL0034 Expression and purification of Protein interactions Bifidobacterium longum NCC2705
CLC: Q935
Type: Master's thesis
Year: 2010
Downloads: 25
Quote: 0
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Abstract


Bifidobacteria within the intestinal tract of humans and animals inhabit the largest number of the most important physiological functions of bacteria, as the core of the micro-ecology research, the host has a biological barrier, nutrition, immunization, control endotoxemia, anti-aging, anti- The physiological role of tumor gene therapy of tumors, but also a very satisfactory guide carrier. Fermentation viable dairy Bifidobacterium industrial production technology bottleneck, international development Bifidobacterium industry key issues that must be addressed. Bifidobacteria in the body nutrition comes mainly from plant material containing fructose-based, proven Bifidobacterium fructose transport and metabolism can reveal its adaptability in the gut, and has a certain theoretical significance and potential applications value. In this study, proteomics and bioinformatics to determine the fructose-induced regulation of fructose transporter protein, through its cloning and expression in vitro experimental study with substrate binding specificity, GST Subsidence fructose transporter system proteins interactions. To obtain the following results: (1) using pH4-7 IPG strips Bifidobacterium protein sample analysis of two-dimensional electrophoresis, a clear distribution of the protein on the gel, the results show that fructose as carbon source ABC transport system in the sugar-binding protein (BL0033) higher expression levels. (2) Construction of the the bl0033 gene expression vector pGEX-4T-1-b10033, successfully transformed into BL21 and expression of the fusion protein GST-BL0033; GST-tagged fusion protein was purified using glutathione-Sepharose 4B resin beads, and found BL0033 combined with fructose specificity. (3) Construction of a the bl0034 gene expression vector pGEX-4T-1-b10034, successfully transformed BL21 and a large number of expression of the fusion protein GST-BL0034; the protein was purified using glutathione-Sepharose 4B resin bead, be able to determine the binding BL0034 ATP molecules. (4) Building bl0033 bl0034 two gene expression vector pET32a-bl0033/bl0034, successfully transformed into BL21 and expression of two proteins; purified with a nickel column. GST settlement method to study the interaction between the BL0033 and BL0034, BL0033 and BL0034 found the interaction. Indicating that Bifidobacterium absorption of fructose may be a special transport system by BL0033 and BL0034 joint action to complete the.

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