In the present study, six commonly used promoters, including cytomegalovirus major immediate-early (CMV), the CMV enhancer fused to the chicken beta-actin promoter (CAG), human elongation factor-1α (HEF-1α), mouse cytomegalovirus (mouse CMV), Chinese hamster elongation factor-1α (CHEF-1α), and phosphoglycerate kinase (PGK), a CMV promoter mutant and a CAG enhancer, were evaluated to determine their effects on transgene expression and stability in transfected CHO cells. The promoters and enhancer were cloned or synthesized, and mutation at C-404 in the CMV promoter was generated; then all elements were transfected into CHO cells. Stably transfected CHO cells were identified via screening under the selection pressure of G418. Flow cytometry, qPCR, and qRT-PCR were used to explore eGFP expression levels, gene copy number, and mRNA expression levels, respectively. Furthermore, the erythropoietin (EPO) gene was used to test the selected strong promoter. Of the six promoters, the CHEF-1α promoter yielded the highest transgene expression levels, whereas the CMV promoter maintained transgene expression more stably during long-term culture of cells. We conclude that CHEF-1α promoter conferred higher level of EPO expression in CHO cells, but the CMV promoter with its high levels of stability performs best in this vector system.

A series of artificial receptors, based on a sulfonamido system, have been designed and synthesized. The interaction of these receptors with biologically important anions was determined by UV-vis, (1)H NMR titration and electrochemical experiments. Results indicate that these receptors show high recognition abilities for fluoride (F(-)) or acetate (AcO(-)), moderate affinities for dihydrogen phosphate (H(2)PO(4)(-)) or hydroxyl (OH(-)) and almost no affinities for chloride (Cl(-)), bromide (Br(-)) or iodide (I(-)). (1)H NMR titration shows that the interaction between the receptors and anions depends on the hydrogen-bond formation. The Co(III)/Co(II) redox signals of receptor and disappear gradually when the fluoride or acetate anions are added. Moreover, visual color changes accompany guest binding, enabling this system to act as colorimetric anion sensors. The colorimetric properties of these sensors are ascribed to the hydrogen-bond formation and the colorimetric group quinoxaline.

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A strategy for the synthesis of isoquinolylselenocyanates and quinolylselenocyanates through electrophilic selenocyanogen cyclization has been developed. The feature of this reaction is that the sequential process was induced directly by generated in situ pseudohalogen (SeCN) 2 generated in situ. Additionally, the obtained selenocyanates allowed functional group diversification, which could be potential intermediates for valuable compounds.

A novel artificial receptor, (3'-nitrobenzo)[2,3-d]-(3''-nitrobenzo)[9,10-d]-1,4,8,11-tetraazacyclotetradecane-5,7,12,14-tetraone, was designed and synthesized. The interactions of this receptor with different anions were determined by UV-vis and (1)H NMR titration experiments, and their affinity constants to different anions were compared with those of other similar/different systems. The results indicated that this receptor showed a high selective and recognitive ability for F(-) among F(-), Cl(-), Br(-), I(-), AcO(-), OH(-), and H(2)PO(4)(-). Moreover, the low energy configuration of this receptor was further determined by means of theoretical investigations.