Under reducing conditions the apparent molecular weight of the synthesized TGF- β1 monomer is about 13 kD. For expression of the recombinant protein, the expression plasmid was expressed in bacterial host strain BL21 (DE3) carrying plasmid pLysS with capacity to express lysozyme. In an attempt to study the structural and functional details of the mature 25 kD-form of TGF- β1, we constructed an expression system for recombinant expression of rat TGF- β1 under control of the T7 RNA polymerase in Escherichia coli. However, there are several notable differences in structure and flexibility which may be related to specialized functions of various TGF- βs 4. The three-dimensional solution structure of the 25 kD disulfide-linked human TGF- β1 homodimer and the crystal structure of TGF- β2 4, 5, 6, 7, 8 reveal that TGF βs are strikingly similar. Although the release from the latent complex is not fully understood it is most likely that highly regulated proteolytic processings are important to unmask and physiologically activate TGF- β1. This cytokine is biological inert when secreted by most tissues in form of the so called latent TGF- β1 complex 2, 3. Five distinct TGF- β isoforms have been cloned from various sources, among them TGF- β1 is the most intensively studied and best characterized one. Transforming growth factor- βs (TGF- βs) are a family of 25 kD- polypeptides with multifunctional actions in controlling the growth, differentiation and function of a broad range of target cells of both epithelial and mesenchymal origin 1. The mature recombinant rat TGF- β1 expressed in this study provides a useful tool for future detailed structural and functional studies. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF- β1 antibody. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. The molecular weight of expressed TGF- β1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. This system allowed an active and selective synthesis of recombinant TGF- β1. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF- β1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. Improved Dual Color Standards are brighter than the competition on both gels and blots.In order to study structure-function details of TGF- β1,the recombinant mature form of rat TGF- β1 was expressed in bacteria. Increased overall brightness provides visual robustness throughout the entire protein electrophoresis and western blotting workflow. 8 blue-stained bands and 2 pink reference bands (25 and 75 kD).Mixture of 10 recombinant proteins of precise molecular weights (10–250 kD).Use improved Precision Plus Protein Dual Color Standards for molecular weight estimation on SDS-PAGE gels and western blots. Place this handy selection guide magnet on your refrigerator or lab bench to eliminate the guesswork and make protein standard selection simple. LC-MS innovation: Improve analytical accuracy Which microbioreactor modules are best for you Which solid support is right for your oligo synthesis?
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