基因工程大肠杆菌肠毒素菌苗研究进展

在传染性腹泻中产肠毒素性大肠杆菌(ETEC)的作用已引起人们的高度重视。DNA重组技术的发展,为研制大肠杆菌肠毒素基因工程菌苗提供了新途径。本文叙述了大肠杆菌肠毒素的结构及其基因分析,介绍了基因工程菌苗的研究近况。     

抗生素产生菌原生质体融合技术的研究进展

原生质体融合技术又称细胞融合技术,是一个人工实验系统,它将遗传性不同的两种细胞融合为一种新细胞。这种在细胞水平上的遗传操作属细胞工程,是现代生物技术的一个重要方面,因该技术用于植物及微生物时,要去掉细胞壁,故称原生质体融合。从该技术现有内容来看,它不再仅仅是原生质体融合,而且已形成了以原生质体为基础的DNA或质粒DNA转化、转导技术,它们是基因工程不可缺少的组成部分。     

基因工程制药应用及研究进展

基因工程制药通过DNA重组技术,进行无性繁殖和基因表达来获得所需核酸或蛋白质,产品主要有细胞因子、蛋白质激素、抗血栓药等,广泛应用于癌症、免疫性疾病、抗病毒、心血管疾病等。本文就基因工程蛋白质药物为主的基因工程制药的概况进行综述。     

微生物基因工程育种技术的研究进展

微生物基因工程育种是20世纪末发展起来的分子定向育种技术,该文重点概述了基因定点突变、易错PCR、DNA重排及基因组重排这几种微生物基因工程育种的基本原理和操作过程,并对这些方法的研究进展进行了扼要综述.     

关于基因工程社会与伦理问题的思考

基因工程,亦即重组DNA分子技术,是七十年代初期才问世的一类新兴生物技术。由于该技术的自身特点,从一开始就引起了一场激烈的争论。本文通过对这场历史争论的反思,探讨了有关基因工程的社会及伦理问题。     

赛诺金治疗病毒性肝炎的心理护理

赛诺金注射剂是采用DNA重组技术生产的高纯度α-1b型基因工程干扰素。是用健康人白细胞来源的干扰素基因克隆和表达的基因工程药物,具有广泛的抗病毒、抗肿瘤及免疫调节功能,可抑制病毒在细胞内复制,增强NK细胞活性,有效地遏制病毒的侵袭。     

高压亲和层析在生物学技术领域的应用

<正> 前言 DNA重组技术或基因工程无疑是二十世纪生物学的一项最伟大成就,从七十年代开始,基因工程在疫苗、多肽、蛋白、激素、干扰素、酶制剂、血液因子等领域的应用开发中取得了十分瞩目的效益,基因工程和杂交瘤等现代生物学技术的产品以及其他生化制品都面临着如何有效的分离提纯问题,即把研究成果转为大规模生产的商品化。生物大分子的经典分离技术,包括多糖基离子交换、分子筛、色谱聚焦等,这些形式各     

生物技术及其在医药工业中的应用前景(上)

现代生物技术包括DNA重组技术、杂交瘤技术和组织培养技术。本文主要对这三种新技术在医药工业中的应用及其发展前景作一概述。一、基本技术及它们的相互关系 DNA重组(recombinant DNA)技术,亦有基因工程(gene engineering)、基因操作(gene manipulation)、基因拼接(genesplicing)、遗传工程(genetic engineering)等名称。实际上这些名称的含义均相同。     

第三代基因工程-代谢工程

代谢工程(Metabolic Engineering),亦称途径工程(Pathway Engineering)和代谢设计(Metabolic Design),是一门利用分子生物学原理系统分析细胞代谢网络、并通过DNA重组技术合理设计细胞代谢途径及遗传修饰,进而完成细胞特性改造的应用性学科.鉴于它涉及到多基因的联合协同表达控制机制,与传统的蛋白多肽单基因表达(第一代基因工程)以及基因定向突变(第二代基因工程)已有显著区别,故作者将之看作为第三代基因工程.     

重组降压肽的设计与发酵生产

本发明应用DNA重组技术，通过基因工程菌发酵法来制备。具体涉及重组降压肽相关基因设计与合成，构建重组降压肽表达载体，用所述表达载体转化宿主构建基因工程菌，利用所述基因工程菌发酵生产重组降压肽。先设计出一条前体肽，该肽可以是一种降压肽的重复体也可是多种降压肽的混合体。例如本发明采用混合肽，LRP，LYPVK，AVNPIR，GHKIATFQER，QVPYPQR。[第一段]     

蛋白质工程技术及其在生物药物研发中的应用

蛋白质工程技术是在基因水平上通过分子设计和DNA重组技术合成自然界不存在的蛋白质的新技术。介绍定点突变、体外定向进化和tRNA介导蛋白质工程3类蛋白质工程技术,并综述其在生物药物研发中的应用。     

利用基因工程技术选育氨甲酰妥布霉素高含量菌种

目的以产生安普霉素、氨甲酰妥布霉素和氨甲酰卡那霉素3个组分的黑暗链霉菌为出发菌株,利用基因工程技术选育氨甲酰妥布霉素含量高的菌种。方法克隆安普霉素生物合成基因,通过基因阻断技术破坏安普霉素生物合成基因,阻断安普霉素生物合成。结果获得了氨甲酰妥布霉素含量高的菌种。结论利用基因工程技术可以有效地改良抗生素组分。     

Matrices and scaffolds for DNA delivery in tissue engineering

Regenerative medicine aims to create functional tissue replacements, typically through creating a controlled environment that promotes and directs the differentiation of stem or progenitor cells, either endogenous or transplanted. Scaffolds serve a central role in many strategies by providing the means to control the local environment. Gene delivery from the scaffold represents a versatile approach to manipulating the local environment for directing cell function. Research at the interface of biomaterials, gene therapy, and drug delivery has identified several design parameters for the vector and the biomaterial scaffold that must be satisfied. Progress has been made towards achieving gene delivery within a tissue engineering scaffold, though the design principles for the materials and vectors that produce efficient delivery require further development. Nevertheless, these advances in obtaining transgene expression with the scaffold have created opportunities to develop greater control of either delivery or expression and to identify the best practices for promoting tissue formation. Strategies to achieve controlled, localized expression within the tissue engineering scaffold will have broad application to the regeneration of many tissues, with great promise for clinical therapies.     

ADAM33基因S2位点突变与COPD患者病程及年龄的相关性研究

目的探讨解整合素—金属蛋白酶33(ADAM33)基因S2位点多态性与慢性阻塞性肺疾病(COPD)发病以及患者病程长短、年龄大小的相关性。方法对84例COPD患者(COPD组)和96例健康者(对照组),应用聚合酶链反应—限制性片段长度多态性(PCR-RFLP)及DNA测序的方法,以对ADAM33基因S2位点多态性与患者病程及年龄进行分析。结果 COPD组ADAM33基因S2位点C/C和C/G基因型频率均高于对照组,差异有统计学意义(P<0.05)。但C/C、C/G、G/G基因型频率与COPD病程长短、年龄大小比较均无统计学意义(P>0.05)。结论 ADAM33基因S2位点单核苷酸多态性(SNPs)与COPD相关;ADAM33SNPsS2突变可能为先天性基因突变。     

制药工程专业开设《中药分离工程》课程的实践

结合国家实施的中药现代化战略,从工程学的角度探讨了在制药工程专业(中药制药工程方向)开设《中药分离工程》课程的重要性及意义,并对制药工程专业开设《中药分离工程》课程进行了实践。     

制药反应工程课程教学改革

制药反应工程是制药工程专业学生的专业基础课,对培养学生的工程概念、强化工程分析能力具有十分重要的作用。该文从制药反应工程的课程特点出发,在教学内容、教学方法、培养学生综合能力等方面进行有益的教改探索。     

制药工程学教材的内容和特点

制药工程学是制药工程专业的主干专业课程，也是我校制药工程专业的主要特色专业课程。作者根据多年来从事制药工程领域教学和科研工作的经验，编著了制药工程学教材，并简要介绍了制药工程学教材的内容和特点。     

对学生工程素质培养的探讨

工程素质的培养是工程类专业教学的核心 ,具有很强的实践性和可操作性。以课程设置改革和实验基地建设两个方面对制药工程专业 (生物制药方向 )学生工程素质培养的必要性进行了阐述 ,认为课程体系工程学科建设和生产中试基地的建设是提高学生工程素质能力和锤炼科研创新能力的支撑平台 ,并在短时间内如何增加学科建设的投入和提高学生动手实践能力提出了自己的看法     

A Fibrin Glue Composition as Carrier for Nucleic Acid Vectors

Purpose

Gene delivery from biomaterials has become an important tool in tissue engineering. The purpose of this study was to generate a gene vector-doted fibrin glue as a versatile injectable implant to be used in gene therapy supported tissue regeneration.

Methods

Copolymer-protected polyethylenimine(PEI)-DNA vectors (COPROGs), naked DNA and PEI-DNA were formulated with the fibrinogen component of the fibrin glue TISSUCOL® and lyophilized. Clotting parameters upon rehydration and thrombin addition were measured, vector release from fibrin clots was determined. Structural characterizations were carried out by electron microscopy. Reporter and growth factor gene delivery to primary keratinocytes and chondrocytes in vitro was examined. Finally,chondrocyte colonized clots were tested for their potency in cartilage regeneration in a osteochondral defect model.

Results

The optimized glue is based on the fibrinogen component of TISSUCOL®, a fibrin glue widely used in the clinics, co-lyophilized with copolymer-protected polyethylenimine(PEI)- DNA vectors (COPROGs). This material, when rehydrated, forms vector-containing clots in situ upon thrombin addition and is suitable to mediate growth factor gene delivery to primary keratinocytes and primary chondrocytes admixed before clotting. Unprotected PEI-DNA in the same setup was comparatively unsuitable for clot formation while naked DNA was ineffective in transfection. Naked DNA was released rapidly from fibrin clots (>70% within the first seven days) in contrast to COPROGs which remained tightly immobilized over extended periods of time (0.29% release per day). Electron microscopy of chondrocytecolonized COPROG-clots revealed avid endocytotic vector uptake. In situ BMP-2 gene transfection and subsequent expression in chondrocytes grown in COPROG clots resulted in the upregulation of alkaline phosphatase expression and increased extracellular matrix formation in vitro. COPROG-fibrinogen preparations with admixed autologous chondrocytes when clotted in situ in osteochondral defects in the patellar grooves of rabbit femura gave rise to luciferase reporter gene expression detectable for two weeks (n=3 animals per group). However, no significant improvement in cartilage formation in osteochondral defects filled with autologous chondrocytes in BMP-2-COPROG clots was achieved in comparison to controls (n=8 animals per group).

Conclusions

COPROGs co-lyophilized with fibrinogen are a simple basis for an injectable fibrin gluebased gene-activated matrix. The preparation can be used is complete analogy to fibrin glue preparations that are used in the clinics. However, further improvements in transgene expression levels and persistence are required to yield cartilage regeneration in the osteochondral defect model chosen in this study.