高等植物叶绿体基因工程

叶绿体基因工程作为一项新技术具有一系列传统核基因工程所不具备的优点,在基础性及应用性研究中极具吸引力,已经成功应用于了解质体基因组,调控植物代谢系统,农作物抗旱、抗虫、抗病、抗除草剂及以植物为生物反应器生产抗体、疫苗等方面的研究。本文主要介绍叶绿体基因工程的原理、操作体系及其在高等植物中的应用。     

用生态工程技术控制农村非点源水污染

非点源污染的本质是农村生态系统的严重失调,生态工程技术是进行流域生态修复,强化物质循环的有效方法,非点源污染的治理策略有控制径流污染和控制源头污染两个方面。本文介绍了多水塘系统,植被缓冲带,湿地系统,生态农业、坡面生态工程和污染物生态处理6种生态治理技术,它们的有机结合可形成控制非点源污染的流域生态工程,执行时,应该因地制宜,还要加强资金、管理和教育的投入。     

Microbial production of propionic acid from propionibacteria: Current state,challenges and perspectives

Propionic acid (PA) is an important building block chemical and finds a variety of applications in organic synthesis, food, feeding stuffs, perfume, paint and pharmaceutical industries. Presently, PA is mainly produced by petrochemical route. With the continuous increase in oil prices, public concern about environmental pollution, and the consumers’ desire for bio-based natural and green ingredients in foods and pharmaceuticals, PA production from propionibacteria has attracted considerable attention, and substantial progresses have been made on microbial PA production. However, production of PA by propionibacteria is facing challenges such as severe inhibition of end-products during cell growth and the formation of by-products (acetic acid and succinic acid). The integration of reverse metabolic engineering and systematic metabolic engineering provides an opportunity to significantly improve the acid tolerance of propionibacteria and reduce the formation of by-products, and makes it feasible to strengthen the commercial competition of biotechnological PA production from propionibacteria to be comparable to the petrochemical route.     

奇异果甜蛋白及其基因工程

奇异果甜蛋白(thaumatin)是迄今为止最甜的物质之一,对其研究具有很重要的意义。奇异果甜蛋白的生化性质基本清楚,基因序列和氨基酸序列都已测定。它的甜味可能是由奇异果甜蛋白上特定基团和受体结合引起的。对奇异果甜蛋白的生理功能知之甚少。近二十年来,在奇异果甜蛋白的基因工程上取得了一定进展,但仍然存在许多困难。 Abstract:Thaumatin is one of the sweetest substances known to date,it is important to study the thaumatin.The biochemical properties of thaumatin have been clarified clearly.Thaumatin had been isolated and sequenced.The mechanism of the sweetness of thaumatin may be due to the combination of some special groups and the receptors.The exact function of thaumatin is still not clear.Although gene engineering of thaumatin has been carried out for 20 years,there are still some difficulties to be solved for using in the market.     

蜘蛛丝蛋白研究进展

由于蜘蛛丝蛋白分子高度重复的一级结构、特殊的溶解特性和分子折叠行为以及具有形成非凡力学特性丝纤维的能力而引人注目。本文从蛛丝蛋白基因、天然蛛丝形成过程、蛛丝蛋白的基因工程生产及蛛丝蛋白的应用前景等几个方面着重介绍了近20年来对蛛丝蛋白的研究进展。围绕蛛丝蛋白展开的研究将有助于揭示蛋白质一级结构、蛋白质分子折叠与蛋白质大分子特性之间的内在联系。     

人新基因HBRP基因工程细胞系的建立

在研究牛精浆蛋白 (bovineseminalplasmaproteins ,BSPproteins)及其相关蛋白在受精卵发育过程中的重要作用时 ,在人睾丸组织中发现并克隆了一个与BSP蛋白相关新基因的序列 ,其开放阅读框架 (openreadingframe ,ORF)编码了一个含 2 2 3个氨基酸残基的蛋白质 ,氨基酸序列中含有4个纤连蛋白Ⅱ型结构域 ,与BSP蛋白在结构上有一定的相似性 ,称其为人BSP相关蛋白 (humanBSP relatedprotein ,HBRP) ,并在GenBank注册 ,登录号为AF2 791 4。预测该蛋白是与BSP蛋白功能相关的结合蛋白 ,并已成功地将该基因定位在 1 9号染色体 1区 3带上。为进一步研究该新基因的生物学功能 ,利用重组质粒pEGFP C2 HBRP转染人胚肾HEK2 93细胞 ,通过G41 8筛选出药物抗性克隆 ,建立了稳定的基因工程细胞系 ,为该基因功能的研究奠定了基础 ,对该基因功能的进一步研究有重要的意义 义。     

人工湿地及其在工业废水处理中的应用

论述了人工湿地污水处理技术的机理和优点,人工湿地利用基质、植物和微生物这个复合生态系统的物理、化学和生物的三重协调作用,通过过滤,吸附、共沉淀、离子交换,植物吸收和微生物分解来实现对废水的高效净化,同时通过营养物质和水分的生物地球化学循环,促进绿色植物生长并使其增产,实现废水的资源化与无害化;人工湿地污水处理系统具有出水水质稳定,对营养物质去除能力强,基建和运行费用低,技术含量低,维护管理方便,耐冲击负荷强,适于处理间歇排放的污水和具有美学价值等优点,该技术不仅能够在发展中国家和发达国家的城市生活污水处理中广泛应用,其在工业废水处理中的应用也正在不断受到重视,根据人工湿地在工业废水处理中的研究和应用现状,指出了人工湿地处理特殊工业废水的前景及今后的研究方向。     

Protein Engineering Allows for Mild Affinity-based Elution of Therapeutic Antibodies

Presented here is an engineered protein domain, based on Protein A, that displays a calcium-dependent binding to antibodies. This protein, Z_Ca, is shown to efficiently function as an affinity ligand for mild purification of antibodies through elution with ethylenediaminetetraacetic acid. Antibodies are commonly used tools in the area of biological sciences and as therapeutics, and the most commonly used approach for antibody purification is based on Protein A using acidic elution. Although this affinity-based method is robust and efficient, the requirement for low pH elution can be detrimental to the protein being purified. By introducing a calcium-binding loop in the Protein A-derived Z domain, it has been re-engineered to provide efficient antibody purification under mild conditions. Through comprehensive analyses of the domain as well as the Z_Ca–Fc complex, the features of this domain are well understood. This novel protein domain provides a very valuable tool for effective and gentle antibody and Fc-fusion protein purification.     

Enhanced production of succinic acid by metabolically engineered<Emphasis Type="Italic">Escherichia coli</Emphasis> with amplified activities of malic enzyme and fumarase

Apfl ldhA double mutantEscherichia coli strain NZN111 was used to produce succinic acid by overexpressing theE. coli malic enzyme gene (sfcA). This strain, however, produced a large amount of malic acid as well as succinic acid. After the analyses of the metabolic pathways, thefumB gene encoding the anaerobic fumarase ofE. coli was co-amplified to solve the problem of malic acid accumulation. A plasmid, pTrcMLFu, was constructed, which contains an artificial operon (sfcA-fumB) under the control of the inducibletrc promoter. From the batch culture of recombinantE. coli NZN111 harboring pTrcMLFu, 7 g/L of succinic acid was produced from 20 g/L of glucose, with no accumulation of malic acid. From the metabolic flux analysis the strain was found under reducing power limiting conditions by severe reorientation of metabolic fluxes.     

人骨髓间充质干细胞向血管内皮细胞诱导分化的研究

目的:分析人骨髓间充质干细胞(hMSCs)和脐静脉内皮细胞(hUVECs)的基因表达差异,探讨体外基因转染诱导内皮分化的可行性以及作为血管组织工程种子细胞来源的应用前景。方法:分别从人骨髓和脐静脉分离间充质干细胞(hMSCs)和内皮细胞(hUVECs),扩增培养后进行流式细胞仪、免疫细胞化学,免疫荧光鉴定和超微结构观察。通过BiostarH-40S表达谱芯片分析,选择两者的差异表达基因,导入hMSCs,经RT-PCR、ELISA鉴定该基因的转染和表达,并分析hMSCs的内皮分化程度。结果:hMSCs表达内皮细胞的多种特异性mRNA,经VEGFl65基因瞬时转染后RT-PCR有明显条带,ELISA定量检测VEGF165蛋白表达为(707.9±11.3)ng/L,同时CD44表达明显下调38.80%,CD31则明显上调达56.82%,FI-1,FVⅢAg和CD34的表达也有不同程度升高。结论:hMSCs具有内皮分化潜能,体外基因转染诱导hMSCs产生功能性内皮细胞和组织工程化血管具有广阔前景。