利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

Agricultural practice changes with cultivating genetically modified herbicide-tolerant oilseed rape

To facilitate weed suppression, oilseed rape or canola (Brassica napus) has been genetically modified (GM) over the last two decades to incorporate herbicide tolerance (HT). The introduction of oilseed rape in commercial agriculture raises concerns about potential adverse agronomic and environmental effects linked to both the genetic modification and altered agricultural practice. How will this new crop modify the existing cropping practice? What are the potential agro-environmental implications of its introduction? This paper analyses and schematises recorded and conceivable agricultural practice changes with GMHT oilseed rape introduction and its likely agro-environmental effects. It develops a conceptual systematics of influencing factors, subsequent practice changes and likely agro-environmental effects. The results can be used to design the mandatory monitoring of adverse GM crop effects.     

巴基斯坦转基因作物发展概况

近年来,发展中国家在转基因技术与商业化发展上取得了巨大进展。巴基斯坦是同中国有着深厚友谊的国家,是“一带一路”政策亚欧非大陆的重要一站。本文介绍了巴基斯坦转基因技术发展现状及转基因商业化情况,研究其管理框架及政策变化等,分析其在政策、监管、知识产权、消费者认知度等方面存在的问题。研究结果对中国转基因政策的制定具有借鉴意义。     

Rapid screening of genetically modified ingredients in soybean and cotton processing by-product and waste using direct qPCR

To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste, direct quantitative PCR (qPCR) kit-Taqman which omitting multi genomic DNA preparing steps was developed in this study. A total of 18 oil crop processing by-products and wastes including 10 soybean and 8 cotton materials were collected from food processing factories. Compared with 2 commercial direct qPCR kits, conditions of DNA releasing procedure and PCR amplification were optimized. Element screening was performed at the initial step of genetically modified (GM) ingredient testing procedure via direct qPCR. GM event identification was carried out in positive samples by initial screening. Totally 5 screening elements (P–35S, T-NOS, Cp4-epsps, bar and pat) for soybean materials and 6 screening elements (P–35S, T-NOS, NPTII, Cry1Ac, bar and pat) for cotton samples were detected. In GM event identification, MON531 and MON1445 were found in cotton materials. Results were further confirmed by real-time PCR with DNA extraction and purification. The direct qPCR system proposed by this research was convenient for rapid screening and identification of GM ingredients in oil crop primary by-product and waste.     

牛冠状病毒4H12/1D4单链基因片段在大肠杆菌中的表达

用ＥｃｏＲＩ酶切ＰＲ５Ｂ４质粒，采用“Ｖ”字形内槽电洗脱法回收约０．８０４Ｋｂ牛冠状病毒基因工程抗体４Ｈ１２／１Ｄ４单链基因片段，并将其插入载体质粒ＰＥＴ－１７ｂ的ＥｃｏＲＩ位点构成重组质粒ＰＥＴ－Ｄ４，将重组质粒转化ＤＨ５α感受态细胞，在ＩＰＴＧ的诱导下表达，细菌裂解物经ＳＤＳ－ＰＡＧＥ电泳筛选，发现含ＰＥＴ－Ｄ４细菌经诱导新增一条约２８ＫＤα蛋白带，该蛋白带与设计基因工程抗体的大小相符，经光密度扫描，表达量约占菌体总蛋白的１４％     

转基因植物潜在的生物入侵问题

通过对转基因植物安全性和外来物种的生物入侵问题的研究发现，转基因植物作为自然界中本来不存在的外来物种，由于其自身的生态幅宽、繁殖能力强等竞争优势，可以通过花粉散布、种子扩散和杂交等途径造成外缘基因的逃逸，从而导致其对生态环境及生物多样性产生一定的影响。     

酸化膨润土对印染废水脱色实验研究

将钙基膨润土酸性活化,然后对印染废水脱色处理,其一次性COD去除率达76%,色度去除率达87%。若经双层滤料柱进行过滤处理,色度去除率达93%以上,COD去除率达78%,处理后的印染废水达到国家规定的排放要求。     

转基因生物技术的生态风险及对策研究

转基因生物技术是现代分子生物技术的一个分支学科,它的开发和应用是人类智慧的产物,人们在利用生物技术造福的同时,也可能带来安全问题,特别是转基因生物技术的滥用对生物多样性、生态环境及人体健康都可能构成危险或潜在的风险,即生态风险问题。各个国家和地区通过制定一系列的法律、法规对转基因生物技术及应用进行监管。我国当前没有关于转基因生物技术生态风险防范的全面而系统的法规体系,在分析和比较国内外转基因生物技术相关立法的基础上,提出相应的完善对策。     

转基因大豆MON89788检测质粒标准分子构建与应用

[目的]构建适用于转基因大豆MON89788检测的质粒标准分子.[方法]利用定性PCR和连接、转化等分子克隆技术,将大豆内标准基因lectin、MON89788的3'端特异性序列和5'端特异性序列依次克隆到pMD18-T载体上,获得质粒标准分子pMD-LM3M5,并进行适用性验证.[结果]获得了3 700 bp的质粒标准分子,其中重组DNA片段1 029 bp.该质粒标准分子的定性PCR检测灵敏度达到10 copy.[结论]该研究构建的质粒标准分子pMD-LM3M5能替代NON89788基体标准品,用于MON89788大豆及其产品的定性PCR检测. Abstract： [Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788.[Method]the lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment.The limits of qualitative detection of the PRM were 10 copies,[Conclusion]The PRM constructed in this study was suitable for the identification of MON89788 event.     

中国转基因作物产业化分析

伴随着经济全球化和世界人口的不断地增长,转基因作物产业化对全球的经济、社会、环境起了重要的影响。中国作为转基因作物种植面积400万hm2的发展中大国,转基因作物产业同样对经济、社会和环境起重要作用。文中对中国转基因作业产业的现状及研究状况进行了分析并提出:在中国转基因作物产业化未来的发展中,要加强转基因作物研发能力;构建以企业为主体,市场为导向,形成和完善生产、加工、销售一体化的转基因作物产业体系;强化转基因作物安全管理,完善生物安全评价体系和转基因作物的科普工作。中国做好转基因作物产业化不仅能保障粮食安全,而且能产生巨大的经济、社会和环境效益,为中国可持续发展战略提供不竭动力。