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酵母表达体系及其在抗体工程中的应用 总被引:6,自引:0,他引:6
酵母是最简单的真核生物,以其为宿主表达异源蛋白具有很大的优越性,近十几年来,各种各样的酵母表达体系陆续发展起来,本文介绍了两种酵母表达系统,并对现有的主要表达体系进行了比较,另外,本文还涉及到酵母在表达抗体或其片段方面的成功应用。 相似文献
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昆虫表达系统是一类应用广泛的真核表达系统,该系统具有与其他高等真核表达系统相似的翻译后修饰、加工及转移外源蛋白的能力。本文介绍了昆虫表达系统的构建过程,并以基因工程抗体为主讨论了外源蛋白在昆虫表达体系中的表达特征。 相似文献
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昆虫表达系统是一类应用广泛的真核表达系统,该系统具有与其他高等真核表达系统相似的翻译后修饰、加工及转移外源蛋白的能力。本文介绍了昆虫表达系统的构建过程,并以基因工程抗体为主讨论了外源蛋白在昆虫表达体系中的表达特征。 相似文献
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人Flt3配体在酵母中的表达及其生物学特性 总被引:6,自引:1,他引:5
Flt3配体(FL)是一种具有促进早期造血功能的细胞因子,能够促进造血干/祖细胞的增殖和分化。为了获得高效表达的人可溶性Flt3配体(rhFL)重组蛋白,采用酵母偏爱的密码子,人工合成rhFL基因,构建表达载体pPICZaA-FL,电转化毕氏酵母9Pichia pastoris),得到稳定分泌rhFL的基因工程菌株,在培养上清中,其表达量达30mg/L。生物学活性检测提示,毕氏酵母系统表达的rhF 相似文献
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t-PA cDNA的克隆及其在毕赤酵母中的表达 总被引:3,自引:0,他引:3
应用PCR方法,在tPAcDNA5′端引入合适的限制酶位点和酵母分泌信号肽,与酵母表达载体pPIC9重组,构建表达质粒pSTEY,利用LiCl转化法转化酵母菌YS108,在MM和MD平板上筛选表型,PCR筛选tPAcDNA与酵母染色体整合而形成的阳性克隆,阳性克隆经甲醇诱导表达后,用SDSPAGE证明表达产物的分子量为6kDa左右,用酪蛋白板溶圈法测定tPA的活性。Muts表型菌表达产物活性最高为2500IU/ml,Westernblot证实表达产物具有天然tPA分子的免疫原性。 相似文献
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The production of recombinant proteins is important in academic research to identify protein functions. Moreover, recombinant enzymes are used in the food and chemical industries, and high-quality proteins are required for diagnostic, therapeutic, and pharmaceutical applications. Though many recombinant proteins are produced by microbial or mammalian cell-based expression systems, plants have been promoted as alternative, cost-effective, scalable, safe, and sustainable expression systems. The development and improvement of transient expression systems have significantly reduced the period of protein production and increased the yield of recombinant proteins in plants. In this review, we consider the importance of plant-based expression systems for recombinant protein production and as genetic engineering tools. 相似文献
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All beetle luciferases have evolved from a common ancestor: they all use ATP, O2, and a common luciferin as substrates. The most studied of these luciferases is that derived from the firefly Photinus pyralis, a beetle in the superfamily of Cantharoidea. The sensitivity with which the activity of this enzyme can be assayed has made it useful in the measurement of minute concentrations of ATP. With the cloning of the cDNA coding this luciferase, it has also found wide application in molecular biology as a reporter gene. We have recently cloned other cDNAs that code for luciferases from the bioluminescent click beetle, Pyrophorus plagiophthalamus, in the superfamily Elateroidea. These newly acquired luciferases are of at least four different types, distinguishable by their ability to emit different colours of bioluminescence ranging from green to orange. Unique properties of these luciferases, especially their emission of multiple colours, may make them additionally useful in applications. 相似文献
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Phage display systems and their applications 总被引:12,自引:0,他引:12
Paschke M 《Applied microbiology and biotechnology》2006,70(1):2-11
Screening phage display libraries of proteins and peptides has, for almost two decades, proven to be a powerful technology
for selecting polypeptides with desired biological and physicochemical properties from huge molecular libraries. The scope
of phage display applications continues to expand. Recent applications and technical improvements driving further developments
in the field of phage display are discussed. 相似文献
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The complexity of biological membranes has motivated the development of a wide variety of simpler model systems whose size, geometry, and composition can be tailored with great precision. Approaches highlighted in this review are illustrated in Figure 1 including vesicles, supported bilayers, and hybrid membrane systems. These have been used to study problems ranging from phase behavior to membrane fusion. Experimental membrane models continue to advance in complexity with respect to architecture, size, and composition, as do computer simulations of their properties and dynamics. Analytical techniques such as imaging secondary ion mass spectrometry have also been developed and refined to give increasing spatial resolution and information content on membrane composition and dynamics. 相似文献
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Members of the Archaea domain are extremely diverse in their adaptation to extreme environments, yet also widespread in "normal" habitats. Altogether, among the best characterized archaeal representatives all mechanisms of gene transfer such as transduction, conjugation, and transformation have been discovered, as briefly reviewed here. For some halophiles and mesophilic methanogens, usable genetic tools were developed for in vivo studies. However, on an individual basis no single organism has evolved into the "E. coli of Archaea" as far as genetics is concerned. Currently, and unfortunately, most of the genome sequences available are those of microorganisms which are either not amenable to gene transfer or not among the most promising candidates for genetic studies. 相似文献
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Verbelen PJ De Schutter DP Delvaux F Verstrepen KJ Delvaux FR 《Biotechnology letters》2006,28(19):1515-1525
In several yeast-related industries, continuous fermentation systems offer important economical advantages in comparison with traditional systems. Fermentation rates are significantly improved, especially when continuous fermentation is combined with cell immobilization techniques to increase the yeast concentration in the fermentor. Hence the technique holds a great promise for the efficient production of fermented beverages, such as beer, wine and cider as well as bio-ethanol. However, there are some important pitfalls, and few industrial-scale continuous systems have been implemented. Here, we first review the various cell immobilization techniques and reactor setups. Then, the impact of immobilization on cell physiology and fermentation performance is discussed. In a last part, we focus on the practical use of continuous fermentation and cell immobilization systems for beer production. 相似文献
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Cell-transistor hybrid systems and their potential applications 总被引:3,自引:0,他引:3
Electrogenic cells fire spontaneous or triggered action potentials (transient changes of their membrane potentials) and can be electronically coupled to external electrodes (arrays). Signals from rat heart-muscle cells were recorded by a field-effect transistor and the results described on the basis of an equivalent circuit. This technique has potential applications in drug screening, such as measuring the dose-response curve of isoproterenol, a beta-adrenergic agonist with a positive chronotropic effect. 相似文献
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Production of serpins using yeast expression systems 总被引:2,自引:0,他引:2
Serpins occupy a unique niche in the field of biology. As more of them are discovered, the need to produce sufficient quantities of each to aid experimental and therapeutic research increases. Yeast expression systems are well suited for the production of recombinant serpins. The genetics of many yeast species is well understood and readily manipulated to induce the targeted over-production of many different serpins. In addition, protease-deficient strains of certain species are available and a few species carry out post-translational modifications resembling those of humans. Yeasts are easy to grow and multiply readily in simple culture media hence the cost of production is low, while the scale of production can be small or large. The disadvantages are the inability of most yeast(s) to perform complex post-translational modifications and a lower product yield of secreted protein compared to intracellular protein production. However, for the intracellular production of serpins, in particular the clade B serpins that do not have complex post-translational modifications, yeast expression systems should be among the first systems considered. 相似文献
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Laccase-mediator systems and their applications: A review 总被引:1,自引:0,他引:1
O. V. Morozova G. P. Shumakovich S. V. Shleev Ya. I. Yaropolov 《Applied Biochemistry and Microbiology》2007,43(5):523-535
The mechanism of operation of laccase-mediator systems (LMSs) in xenobiotic degradation mediated by “true” redox mediators and laccase enhancing agents is considered. Structural formulae of most common laccase mediators and compounds that can be used as agents enhancing the enzyme operation are presented. Examples of LMS application in biotechnology are described. 相似文献