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Disclosed herein are systems and methods for producing recombinant proteins utilizing mutant E. coli strains containing expression vectors carrying nucleic acids encoding the proteins, and secretory signal sequences to direct the secretion of the proteins to the culture medium. Host cells transformed with the expression vectors are also provided.
Systems and methods for the production of malonate in recombinant host cells.
Systems and methods for the production of malonate in recombinant host cells.
Systems and methods for the production of malonate in recombinant host cells.
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Calmodulin (CaM) was used as an affinity tail to facilitate the purification of the green fluorescent protein (GFP), which was used as a model target protein. The protein GFP was fused to the C-terminus of CaM, and a factor Xa cleavage site was introduced between the two proteins. A CaM-GFP fusion protein was expressed in E. coli and purified on a phenothiazine-derivatized silica column. CaM binds to the phenothiazine on the column in a Ca(2+)-dependent fashion and it was, therefore, used as an affinity tail for the purification of GFP. The fusion protein bound to the affinity column was then subjected to a proteolytic digestion with factor Xa. Pure GFP was eluted with a Ca(2+)-containing buffer, while CaM was eluted later with a buffer containing the Ca(2+)-chelating agent EGTA. The purity of the isolated GFP was verified by SDS-PAGE, and the fluorescence properties of the purified GFP were characterized.
With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, beta-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system. Copyright 2002 Elsevier Science (USA).
Abstract All‐perovskite tandem solar cells show great potential to enable the highest performance at reasonable costs for a viable market entry in the near future. In particular, wide‐bandgap (WBG) perovskites with higher open‐circuit voltage ( V OC ) are essential to further improve the tandem solar cells’ performance. Here, a new 1.8 eV bandgap triple‐halide perovskite composition in conjunction with a piperazinium iodide (PI) surface treatment is developed. With structural analysis, it is found that the PI modifies the surface through a reduction of excess lead iodide in the perovskite and additionally penetrates the bulk. Constant light‐induced magneto‐transport measurements are applied to separately resolve charge carrier properties of electrons and holes. These measurements reveal a reduced deep trap state density, and improved steady‐state carrier lifetime (factor 2.6) and diffusion lengths (factor 1.6). As a result, WBG PSCs achieve 1.36 V V OC , reaching 90% of the radiative limit. Combined with a 1.26 eV narrow bandgap (NBG) perovskite with a rubidium iodide additive, this enables a tandem cell with a certified scan efficiency of 27.5%.
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We describe the properties and carrier dynamics of surface InP quantum dots (QDs) on In{sub 0.48}Ga{sub 0.52}P, lattice-matched to GaAs (100). The structures were grown using gas-source molecular beam epitaxy. The average height and lateral size of the dots are in the range of 2–6 and 30–50 nm, respectively. The photoluminescence of the surface dots peaks between 750 and 830 nm, depending on the growth conditions, and is red-shifted compared to the emission of the capped QDs grown under similar conditions. The integrated photoluminescence intensity is comparable to that of the capped QDs. The decay time of both surface and capped QDs is around 1 ns at 15 K. The strong luminescence of surface QDs is explained by the effect of acting vacuum/air as an effective barrier and saturated surface states. Enhancement of the QDs luminescence is observed for the samples coated with a fluorescent dye.
We report magnetoconductance measurements on polymer:fullerene photovoltaic devices in the regime of high fields and low temperatures, where spin polarization of injected carriers plays an important role. The current either decreases or increases with magnetic field, depending on whether the interfacial charge-transfer state lies above or below the intramolecular triplet state in energy. Rapid transitions from triplet charge-transfer states to lower-lying triplet excitons constitute an important loss mechanism that is responsible for the negative magnetoconductance observed.
Two kinds of green InGaN light emitting diodes (LEDs) have been investigated in order to understand the different slopes in logarithmic light output power-current (L-I) curves. Through the analysis of the carrier rate equation and by considering the carrier density-dependent the injection efficiency into quantum wells, the slopes of the logarithmic L-I curves can be more rigorously understood. The low current level, two as the tunneling current is initially dominant. The high current level beyond the peak of the external quantum efficiency (EQE) diminishes below one as the carrier overflow becomes dominant. In addition, the normalized carrier injection efficiency can be obtained by analyzing the slopes of the logarithmic L-I curves. The carrier injection efficiency decreases after the EQE peak of the InGaN LEDs, determined from the analysis of the slopes of the logarithmic L-I curves.