动物乳腺生物反应器的现状和趋势

利用转基因家畜的乳腺生产人类重组蛋白,可以高效获得安全、足量的药用蛋白。本文针对乳腺生物反应器的成功研制,从目的基因的选择、载体构建、转基因技术等方面探讨了动物乳腺生物反应器的研究现状。分析了提高转基因效率和外源蛋白表达水平的技术途径,提出了降低总体成本的战略措施。特别探讨了利用Cre-loxP系统发展“体细胞打靶体细胞核移植技术体系”,高效生产乳腺生物反应器动物的可能性。     

转基因动物乳腺生物反应器与基因工程药物

利用转基因动物-乳腺生物反应器生产药物蛋白,好比在动物身上建“药厂”,可以从动物乳汁中源源不断地获得具有稳定生物活性的基因产品。这是一种全新的药物生产模式,具有投资成本低,药物研制周期短和经济效益高等优点。它已经成为生物技术领域发展的重要方向。本文系统地分析了利用转基因动物乳腺生物反应器生产基因工程药物的优越性、基本生产方法、存在的问题和原因及产业化进展。     

动物乳腺生物反应器的研究进展

利用转基因动物反应器生产药用蛋白是生物技术领域的一次革命,而通过动物乳腺是最好的渠道,药物蛋白可通过转基因动物的乳腺随乳汁分泌出来,产量高、易纯化,因此乳腺类似于一个制药工厂。国外已有多种蛋白通过乳朱反应器来制备,有些已进入临床研究阶段。本文主要介绍乳腺生物反应器的研究进展情况。     

动物乳腺生物反应器的应用与展望

应用转基因技术生产乳腺生物反应器可以获得高效、安全、足量的人类重组蛋白、药用蛋白及其目的蛋白.本文概述了制备转基因动物过程中目的基因选择、载体构建等技术环节的研究现状;通过叙述乳腺生物反应器的原理和应用现状来分析乳腺生物反应器的优势及特点,为乳腺生物反应器的发展和应用奠定理论基础.     

动物乳腺生物反应器与生物制药

利用动物乳腺生物反应器生产人类所需的药用蛋白,在制药领域展现出了广阔前景。综述了应用动物乳腺生物反应器制药的优越性和应用进展。     

利用动物乳腺生物反应器生产药用蛋白

动物乳腺生物反应器是利用动物乳腺特异性启动子调控元件指导外源基因在乳腺中特异性表达,并从转基因动物奶液中获取重组蛋白。应用动物乳腺生物反应器生产药用蛋白具有生产方式简单,产量大,蛋白能进行翻译后修饰等优点,是具有广阔前景的生物医药产业。本文仅就动物乳腺生物反应器的建立、检测、目的蛋白的分离纯化以及存在的问题等作一综述。     

乳腺生物反应器的研究现状

乳腺生物反应器是将外源基因在哺乳动物的乳腺中特异表达,以转基因动物的乳腺组织生产药用蛋白。采用乳腺生物反尖器生产药用蛋白质是一种全新的生产模式,它已经成为生物技术领域发展的重要方面。乳腺生物反应器具有能够生产出具有完全生物活性的药用收白,纯化简单,投资少,成本低,对环境没有任何污染等优点,转基因动物生产药用蛋白可以获得巨额经济效益。     

转基因技术在制备动物乳腺生物反应器中的应用和发展

利用乳腺生物反应器可以高效获得安全、足量的药用蛋白,在制药工业中具有广阔的应用前景。但是,目前采用的转基因技术由于其各自固有的局限性,未能使乳腺生物反应器的研究取得长足的进步。基因打靶技术和核移植技术的发展为乳腺生物反应器的开发注入了新的活力。本文综合近年来国内外文献,阐述了各种转基因技术的优点与缺陷,同时说明了构建“体细胞打靶-克隆技术体系”在制备大动物的乳腺生物反应器中的必要性。     

转基因动物乳腺生物反应器相关技术及研究进展

转基因动物乳腺生产人类重组蛋白人血清白蛋白(h SA)、人纤维蛋白原(h FIB)、人蛋白C(h PC)、人凝血因子(h F-Ⅷ)和(h F-Ⅸ)等具有较高市场潜力,使其在医学领域获得广泛应用。对转基因动物乳腺生物反应器的优势、目的基因选择、载体构建、基因工程技术、重组蛋白在肿瘤治疗上的应用、当前困境和未来前景等方面进行较为全面的综述,以期能有助于转基因动物生物反应器的研究。此外较详细地探讨了利用CRISPR-Cas基因打靶技术生产高表达量的人类重组蛋白的可能性。     

乳腺生物反应器的研究现状和产业化前景

转基因动物乳腺生物反应器是伴随转基因动物技术而发展起来的一项高新生物技术。利用这项技术,我们可以从动物乳汁中源源不断地获取用于医疗或保健目的的有生物活性的基因产物。这是一种全新的蛋白质生产模式,它将成为许多国家的重要支柱产业之一。本文介绍了乳腺生物反应器的基本概念和基本原理;概述了制备过程中的目的基因选择、载体构建、转基因等技术环节的研究现状;分析了乳腺生物反应器的优势及存在的问题,并就其产业化前景进行了展望。     

Making recombinant proteins in animals--different systems,different applications

Transgenic animal bioreactors represent a powerful tool to address the growing need for therapeutic recombinant proteins. The ability of transgenic animals to produce complex, biologically active recombinant proteins in an efficient and economic manner has stimulated a great deal of interest in this area. As a result, genetically modified animals of several species, expressing foreign proteins in various tissues, are currently being developed. However, the generation of transgenic animals is a cumbersome process and remains problematic in the application of this technology. The advantages and disadvantages of different transgenic systems in relation to other bioreactor systems are discussed.     

Microalgae as platforms for production of recombinant proteins and valuable compounds: progress and prospects

Over the last few years microalgae have gained increasing interest as a natural source of valuable compounds and as bioreactors for recombinant protein production. Natural high-value compounds including pigments, long-chain polyunsaturated fatty acids, and polysaccharides, which have a wide range of applications in the food, feed, cosmetics, and pharmaceutical industries, are currently produced with nontransgenic microalgae. However, transgenic microalgae can be used as bioreactors for the production of therapeutic and industrially relevant recombinant proteins. This technology shows great promise to simplify the production process and significantly decrease the production costs. To date, a variety of recombinant proteins have been produced experimentally from the nuclear or chloroplast genome of transgenic Chlamydomonas reinhardtii. These include monoclonal antibodies, vaccines, hormones, pharmaceutical proteins, and others. In this review, we outline recent progress in the production of recombinant proteins with transgenic microalgae as bioreactors, methods for genetic transformation of microalgae, and strategies for highly efficient expression of heterologous genes. In particular, we highlight the importance of maximizing the value of transgenic microalgae through producing recombinant proteins together with recovery of natural high-value compounds. Finally, we outline some important issues that need to be addressed before commercial-scale production of high-value recombinant proteins and compounds from transgenic microalgae can be realized.     

Developments in transgenic technology: applications for medicine

Recent advances in the efficiency of transgenic technology have important implications for medicine. The production of therapeutic proteins from animal bioreactors is well established and the first products are close to market. The genetic modification of pigs to improve their suitability as organ donors for xenotransplantation has been initiated, but many challenges remain. The use of transgenesis, in combination with the method of RNA interference to knock down gene expression, has been proposed as a method for making animals resistant to viral diseases, which could reduce the likelihood of transmission to humans. Here, the latest developments in transgenic technology and their applications relevant to medicine and human health will be discussed.     

Selection of in vitro produced, transgenic embryos by nested PCR for efficient production of transgenic goats

The production of valuable pharmaceutical proteins using transgenic animals as bioreactors has become one of the goals of biotechnology. However, the efficiency of producing transgenic animals by means of pronuclear microinjection is low. This may be attributed in part to the low integration rate of foreign DNA. Therefore, a large number of recipients are required to produce transgenic animals. We recently developed a transgenic procedure that combined the techniques of goat oocyte in vitro maturation (IVM), in vitro fertilization (IVF), microinjection, preimplantation selection of the transgenic embryos with nested PCR and transferring the transgenic embryos into the recipient goat uterus to produce transgenic goats. Thirty-seven transgenic embryos determined by nested PCR were transferred to thirty-two recipient goats. In the end, four live-born kids were produced. As predicted, all the live kids were transgenic as identified by PCR as well as Southern blot hybridization, The integration rate was 100% (4/4) which was completely in accordance with the results of embryo preimplantation detection. The results showed a significant decrease in the number of recipients required as only 8 recipients (32/4) were needed to obtain one live transgenic goat. We suggest that the transgenic system described herein may provide an improved way to efficiently produce transgenic goats on a large scale.     

转基因动物在输血医学中的应用

转基因动物技术是在动物整体水平研究和表达目的基因的生物技术,其基本特点是：分子及细胞水平操作,组织及整体水平表达,是常规分子生物学理论和技术的拓展和延伸,也是现代生物高技术研究和开发的热点之一。本简述了转基因动物在输血医学领域的应用及其发展前景,包括利用转基因动物生物反应器制备人血浆蛋白和人血红蛋白、建立血传播病毒的感染模型和血液相关遗传模型以及转基因动物与输血医学的基础研究等。     

High-level expressing YAC vector for transgenic animal bioreactors

The position effect is one major problem in the production of transgenic animals as mammary gland bioreactors. In the present study, we introduced the human growth hormone (hGH) gene into 210-kb human alpha-lactalbumin position-independent YAC vectors using homologous recombination and produced transgenic rats via microinjection of YAC DNA into rat embryos. The efficiency of producing transgenic rats with the YAC vector DNA was the same as that using plasmid constructs. All analyzed transgenic rats had one copy of the transgene and produced milk containing a high level of hGH (0.25-8.9 mg/ml). In transgenic rats with the YAC vector in which the human alpha-lactalbumin gene was replaced with the hGH gene, tissue specificity of hGH mRNA was the same as that of the endogenous rat alpha-lactalbumin gene. Thus, the 210-kb human alpha-lactalbumin YAC is a useful vector for high-level expression of foreign genes in the milk of transgenic animals.     

转基因动物乳腺生物反应器的发展概况及人工染色体在其中的应用

转基因动物乳腺生物反应器位点效应的影响是制备转基因动物乳腺生物反应器过程中的主要问题。酵母人工染色体（YAC）和细菌人工染色体（BAC）具有容量大的特性,可以将乳蛋白的整个基因座包括所有调控序列全部装载进去,有可能克服位点效应的影响,是一种理想的载体。YAC和BAC载体转基因技术可能成为避开基因打靶获得高效表达的转基因动物乳腺生物反应器的另一途径.     

Development of patatin knockdown potato tubers using RNA interference (RNAi) technology,for the production of human-therapeutic glycoproteins

Background  

Patatins encoded by a multi-gene family are one of the major storage glycoproteins in potato tubers. Potato tubers have recently emerged as bioreactors for the production of human therapeutic glycoproteins (vaccines). Increasing the yield of recombinant proteins, targeting the produced proteins to specific cellular compartments, and diminishing expensive protein purification steps are important research goals in plant biotechnology. In the present study, potato patatins were eliminated almost completely via RNA interference (RNAi) technology to develop potato tubers as a more efficient protein expression system. The gene silencing effect of patatins in the transgenic potato plants was examined at individual isoform levels.