Efficient Expression and Purification of Fc-fragment-binding Domain and Its Application to Immunoglobulin G Purification

The number of therapeutic monoclonal antibodies used in clinical trials has recently increased dramatically, leading to the development of optimized downstream purification processes[1]. Staphylococcal protein A （SPA）, a cell-wall protein of Staphylococcus aureus, has been developed as a universal ligand for immunoglobulin G （IgG） purification because it binds specifically to the Fc portion of the IgG molecule of many mammals[2]. However, certain characteristics of SPA severely restrict the advancement of the antibody industry.

Efficient expression of green fluorescent protein (GFP) mediated by a chimeric promoter in Chlamydomonas reinhardtii

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii.

EFFICIENT BIT ERROR PROBABILITY EXPRESSION AND VERY TIGHT BOUND FOR MAXIMAL RATIO COMBINING DIVERSITY SYSTEMS OVER RAYLEIGH FADING CHANNEL

Bit Error Probability (BEP) provides a fundamental performance measure for wireless diversity systems. This paper presents two new exact BEP expressions for Maximal Ratio Combining (MRC) diversity systems. One BEP expression takes a closed form, while the other is derived by treating the squared-sum of Rayleigh random variables as an Erlang variable. Due to the fact that the extant bounds are loose and could not properly characterize the error performance of MRC diversity systems, this paper presents a very tight bound. The numerical analysis shows that the new derived BEP expressions coincide with the extant expressions, and that the new approximation tightly bounds the accurate BEP.

Advances in molecular modification and efficient expression of key enzymes for functional saccharide preparation

Functional saccharide is a general term that is often used to refer to the functional oligosaccharides,functional saccharide alcohols,and functional dietary fibers.These functional saccharides exhibit some health benefiting effects,such as having low calorie,preventing dental caries,and regulating intestinal disorders.Functional saccharides are widely used in food,health products,and the healthcare fields.The preparation of functional saccharides is accomplished mainly through reactions involving transglycosylation,isomerization,or hydrolysis catalyzed by glycosyltransferases,saccharide isomerases,and glycohydrolases,respectively.However,the poor catalytic properties of natural enzymes and low fermentation yields have restricted the large-scale industrial production of functional saccharides.Therefore,molecular modification and efficient expression of key enzymes for functional-saccharide preparation are very important for promoting the low-cost large-scale production of functional saccharides.In this report,the recent advances in functional optimization and expression preparation of enzymes related to functional saccharides are reviewed.

Evaluation of parameters affecting Agrobacterium-mediated transient expression in citrus

Agrobacterium-mediated transient expression assays are a convenient alternative to stable expression because they are simple,easy to perform,and achieve gene expression rapidly.This study investigated the factors affecting transient gene expression efficiency in citrus by observing the cryo-sectioning of leaf samples under a laser confocal microscope.These factors included the composition of the infiltration buffer,the Agrobacterium cell density,the leaf development stage,the incubation temperature,and plant genotype.The highest transient expression level of yellow fluorescent protein（YFP）was detected in Mexican lime（Citrus aurantifolia）on the third day after the intermediate-aged leaves were infiltrated with the improved infiltration buffer 1（15 mmol L^-1 2-（N-morpholino）ethanesulfonic acid,10 mmol L^-1 MgCl2,and 200μmol L^-1acetosyringone）,which had an optical density of 0.8 and was incubated at 22°C.Additionally,this transient expression assay was applied to other citrus genotypes.Of note,trifoliate orange（Poncirus trifoliata）and kumquat（Fortunella obovate）had higher expression efficiency than other six genotypes of the Citrus genus.Our study provides research basis for the selection of optimization strategies in transient gene expression and improves the method for available genome investigation in citrus.

Construction of High Level Expression Cell Lines of Erythropoietin

Human erythropoietin gene was ligated into the mammalian high efficiency expression vector pSV2 dhfr by standard procedures of filling in, trimming, ligation and transformation to construct a high efficiency expression vector. After the expression vector pSEPO25 was transfected into CHO dhfr - cells, the cell line which could express high levels of EPO was successfully selected. The result lays the foundation for production of EPO by genetic biotechnology.