多重实时荧光定量PCR分析转基因烟草外源基因拷贝数

[目的]采用多重实时荧光定量PCR方法在一个反应内同时检测转基因烟草中插入外源基因35S、NOS、NPT Ⅱ的拷贝数。[方法]将转基因阳性参照烟草基因组DNA经梯度稀释,通过多重荧光定量PCR反应同时获得烟草内源参照基因NR、外源基因35S、NOS、NPT Ⅱ的相关性标准曲线方程,将待测株系外源基因的Ct值代入标准曲线方程后估算其拷贝数。[结果]获得了上述4个基因的标准曲线,其R2均接近于1,相关性较高。在检测的六个转基因株系中初筛到3株3个外源基因均为单拷贝的株系。[结论]可使用该方法对转基因烟草外源基因插入拷贝数进行估算,为获得稳定遗传材料提供初筛依据。      

转基因生物食品安全检测分析

综合运用当前国际上先进的基于核酸水平的外源转基因成分检测技术,通过DNA提取、Southern杂交试验、转基因结构的检测、外源基因拷贝数的检测等4项试验,从分子水平和生产角度上对转基因肉鸡进行了一次完整的安全检测分析,这一整套方法对生物食品的生产安全性进行了检测,取得了一次的成效。     

常见食用菌中转基因成分定性PCR检测方法的建立

将食用菌转基因研究用的质粒载体(p301-bG1)DNA,添加到19种常见食用菌样品中,作为模拟阳性样品,从中提取出DNA用于PCR分析,建立了常见食用菌中3个大小分别为165、398、599bp的外源基因(NOS、BAR、GUS)的特异性DNA片段的定性PCR检测方法,还进行了二重与三重PCR分析,并通过不同的模板DNA浓度对PCR扩增结果的影响,分析了PCR检测的灵敏度。     

多重PCR 检测转基因水稻的转基因成分

以水稻内源基因SPS、外源抗虫基因Cry1Ab、外源抗虫基因Cry1Ab/Ac、外源抗虫基因Btc、报告基因GUS、NOS终止子和CaMV35S启动子为检测对象,设计7对引物,通过优化PCR扩增体系中不同引物浓度的配比及退火温度,建立水稻转基因成分的七重PCR检测体系。结果表明：建立的七重PCR体系能有效检测出水稻及其他作物(大豆、玉米、棉花籽、菜籽粕)中的转基因成分,检测过程简便、特异性好。     

转基因"华番一号"番茄的筛选和特异性的定性、定量PCR检测方法

为给转基因植物监测提供技术支持，建立了转基因“华番一号”番茄筛选和特异性的定性、定量PCR检测方法。转基因“华番一号”的筛选PCR检测主要以转基因通用元件CaMV35S启动子和NOS终止子为目的基因片段，特异性PCR检测以转基因外源重组子的CaMV35S启动子和反义EFE基因的相邻序列为目的片段；实验同时设立番茄的LAT52基因为转基因番茄定性、定量PCR检测的内对照基因。在所建立的PCR检测体系中，定性PCR筛选和特异性检测的检测极限为68个拷贝，实时定量PCR方法的检测极限为3个拷贝；筛选定量．PCR检测的定量极限为3个拷贝，特异性定量PCR检测的定量极限为25个拷贝。最后通过对2个已知含量的转基因番茄“华番一号”混合试样的检测，证明了该体系可以有效地用于转基因番茄“华番一号”的筛选和特异性的定性、定量PCR检测。     

转基因水稻潮霉素标记基因PCR检测方法的建立及其在基因水平转移研究中的应用

为建立用于基因水平转移研究, 尤其是DNA经加工和消化后稳定性研究的针对转基因水稻潮霉素标记基因hpt（hygromycin phosphotransferase）的定性和实时定量PCR体系,设计针对hpt的上游通用引物多个片段定性PCR扩增体系,以植物叶绿体基因rbcl为内对照,PCR扩增产物经测序验证.将定性PCR中最小片段（236 bp）连接到质粒载体pUC18-pMD T载体上,提取质粒经验证后做外标.应用TaqMan-MGB荧光探针和引物,建立定量的外标校正曲线法,并评价方法的精密度.建立的定性PCR体系能稳定扩增出236 bp～910 bp不同大小的5个hpt片段,并经测序验证.实时定量PCR的线性范围为105～10拷贝（R^2=0.998）,最低能检出10拷贝,重复性好.本研究已成功建立了用于转基因水稻标记基因hpt基因水平转移研究的定性和定量PCR系统。     

转基因苜蓿草J101品系特异性定性PCR检测 方法的建立

目的建立转基因苜蓿草J101品系特异性定性PCR检测方法。方法根据转基因苜蓿草品系J101 5’端外源插入片段与苜蓿草基因组DNA之间的邻接区序列设计引物,建立了转基因苜蓿草J101品系特异性定性PCR检测方法,并对本方法的特异性、灵敏度进行了测定。结果建立的检测方法特异于转基因苜蓿草J101检测,检测最低DNA浓度为(1imit of detection,LOD)为80 pg,相当于50拷贝转基因苜蓿草J101基因组DNA。结论本研究建立的转基因苜蓿草J101品系特异性定性PCR检测方法特异性好,灵敏度高,能够快速、准确地对转基因苜蓿草J101进行检测分析。     

转基因水稻Bt63的外源基因相对含量分析

为分析转基因Bt63稻米外源基因含量,利用新型、灵敏和高通量的实时荧光定量PCR技术,以RBE-4作为转基因水稻的内源参照基因,通过梯度稀释法,分别获得了Bt 63和RBE-4基因的Ct值与起始模板相关性的标准曲线,相关系数分别为0.996 6和0.995 4。通过转基因水稻外源基因(Bt63)和内标基因(RBE-4)起始模板数的比较,测定了外源基因在转基因水稻中的相对含量,这项研究将为建立转基因水稻抗虫能力评价体系和转基因稻谷的流通和监管提供技术支撑。     

叶片衰老诱导FLP重组酶删除转基因烟草外源基因的研究

  目的   “Gene-deletor”系统可实现转基因植物中外源基因的清除。为了研究该系统在转基因烟草中对清除外源基因的作用并最终创制不含外源基因的烟草新种质。  方法   以含有“Gene-deletor”系统的转基因烟株为材料,分析外源筛合报告选融基因Bar::GUS、重组酶基因FLP在不同叶龄叶片中的表达水平,同时通过观察转基因烟草花粉中GUS蛋白的表达活性,分析外源基因GUS在花粉中的删除情况。  结果   （1）相同叶龄不同转基因烟株叶片均表现出GUS活性和对除草剂草铵膦抗性,且3个转基因烟草株系中GUS活性和对除草剂抗性都表现出随着叶龄增大而降低的特点。（2）Real-time PCR和RT-PCR结果进一步证明外源Bar::GUS基因表达水平随叶片发育成熟持续下降,而外源FLP基因的表达水平则出现先增后降的变化趋势,在测定后期两者的表达量均达到最低,（3）花粉GUS组织染色结果表明,“明,色结果降的变化趋势,特系统诱导各转基因植株中花粉外源基因清除效率不同,平均清除率分别78.3%,54.7%和75.2%。  结论   在转基因烟草中,叶片衰老特异表达基因SAG12基因启动子驱动“动启动子驱动达基因,平均清系统（LoxP/FRT）的表达不仅可引起转基因植株叶片中外源基因的特异性清除,还能诱导花粉中外源基因的删除,该技术可为进一步创制培育不含外源基因的烟草新种质提供参考。      

植物源食品DNA制备方法的比较研究

目的：探索可用作PCR方法检测和鉴定植物源转基因食品模板的DNA抽提方法。方法：分别用改良CTAB法、SDS法制备黄豆、玉米、土豆和番茄及其加工产品的DNA，PCR扩增叶绿体基因片段及黄豆、玉米的内参照基因lectin和zein10。结果：改良CTAB法所得DNA作为PCR扩增模板，叶绿体基因片段及lectin和zein10均呈阳性，SDS法所得DNA作为模板时，部分样品内参照基因lectin或zein10扩增阴性。结论：PCR方法检测转基因食品时，应用改良CTAB法制备DNA模板。     

Review of techniques to manufacture micro-hydrogel particles for the food industry and their applications

Microgels are ‘soft’ microscopic cross-linked polymeric particles that are being increasingly exploited in a variety of industries for rheology control, encapsulation and targeted delivery. They are valued because of the ability to tune their functionality to address specific applications in oil recovery, coatings, drug delivery, cosmetics, personal care and foods. Food microgels are typically biopolymer hydrogels in the form of microspheres, nanospheres (also called nanogels), spheroids and fibres. The utilisation of engineered microgels in foods has so far been limited, despite their great potential to address several needs in the food industry, including: satiety control, encapsulation of phytonutrients and prebiotics, texture control for healthier food formulations (e.g. reduced fat products), and targeting delivery to specific areas in the digestive tract. We review the scientific and patent literature on the utilisation and manufacturing methods for producing microgels with an emphasis on micro-hydrogels for food applications.     

12—A CRITIQUE OF SOME HYPOTHESES RELATING TO THE HETEROGENEITY OF THE HIGH-SULPHUR PROTEINS OF WOOL

Joubert and Burns prepared a large number of fractions from the high-sulphur proteins of wool and estimated their molecular weights and amino-acid compositions. Their data have been re-examined in order to look for statistically significant interrelations between amino acids and between the proportion of various amino acids and molecular weight. Statistical analysis of the data is also used to examine the credibility of some hypotheses concerning the mechanism of keratin biosynthesis and to provide further evidence for the existence of families of proteins within the high-sulphur fractions of wool.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of     

The Latest Data about Development of Chinese Printing Industry in 2013

In the 2014 China（Shanghai）International Printing Week,Director Wang Yanbin released the latest data about development of Chinese printing industry in 2013.According to statistics,in 2013,the total output value of Chinese printing industry exceeded 1trillion Yuan for the first time,reaching 1.03985 trillion Yuan.There were 105,000 printing enterprises in China,employees were 3.415 million.The total asset was 1.06247 trillion Yuan;     

The State Council Issued "Directory of Government Approved Investment Projects(2013 Edition)"

正On December 2nd,2013,the State Council issued the notification of"Directory of Government Approved Investment Projects(2013 Edition)"(hereafter referred to as"notification").It is pointed out in the"notification"that in order to further deepen reforms in investment systems and administrative examination and approval systems,simplify administrative procedures and delegate powers to lower levels,earnestly     

Top Ten News of China's Paper Industry 2013

On December 24th, 2013, the meeting on the selection of top 10 news of China＇s paper industry 2013 sponsored by 〈China Paper Newsletters〉 was held in Beijing. The yearly selection of the top l0 news, which began in 2000, has become a brand activity widely recognized in the industry thanks to the support from the authorities at all levels and public participation.     

National Development and Reform Commission Issued Announcement for Selection of Major Preliminary Research Projects for the "13~(th) Five-Year Plan"

In Apri Commi major p Plan＂ （h 2014, the National Development and Reform ssion issued an announcement for selection of reliminary research projects for the ＂13th Five-Year ereafter referred to as ＂The Announcement＂）     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the latest scientific     

China textile machinery: taking off in progress

正Among the 1600 exhibitors who take apart in the ITMA ASIA+CITME2014 2/3 are Chinese manufactures.If the numerous figures failed to attract your attention,the increase of quality should draw your focus.To adopt the demand of developing textile machine market,domestic textile machinery enterprises now follow the slogan of"technology drives development"to enhance product competitiveness.Our domestic sellers will showcase product ranging from spinning,weaving,dyeing and printing,     

Anti-mutagenic compounds from corn

In previous studies with aflatoxin-contaminated corn an uncharacteristic response for AFB₁ in the Salmonella/microsomal mutagenicity assay (Ames test) was observed and the presence of anti-aflatoxin factors in the corn was suggested. In the current study, corn was extracted and fractionated using thin layer chromatography (TLC) using different developing solvent systems and the Ames test was used to monitor for anti-mutagenic activity in the corn fractions. Both Salmonella tester strains TA98 and TA100 with metabolic activation (S9) were used. Several corn fractions, at different stages in the isolation and purification process, showed anti-mutagenic dose-responses when exposed to pure AFB₁. Corn extracts were non-toxic to the tester strains and TLC fractions that showed the best anti-mutagenic dose-responses were selected for further partial characterization analyses.