基因重组技术在工业微生物菌种选育中应用的研究进展

全基因组改组(Crenome shuffling)技术作为一种新型的菌种选育方法,与常见的育种方法相比,具有快速有效、简单易行和实用性强等特点,近年来不断被应用于各种工业微生物菌种的改良研究中.文中介绍了Genome shuffling技术在一系列菌种改良实例中应用现状,为该技术进一步的推广应用莫定基础.     

基因组改组:几株同源酱油曲霉的多亲株电融合育种

酱油酿造中原料的全氮利用率是一个主要的生产指标，通过育种提高生产菌株的中性蛋白酶活是一个有效的方法。利用一株用曲精中分离、纯化的酱油曲霉，制备其分生孢子的原生质体，并优化了原生质体制备的条件。将分生孢子的原生质体进行紫外诱变，得到7株酶活提高幅度较大的紫外诱变株U，以此作为候选株文库，采用基因组改组（Genome shuffling），进行多亲株灭活电融合，获得9株酶活进一步提高的菌株。     

GENOME SHUFFLING技术优化及其应用进展

Genomeshuffling作为一种新型的育种方法,与传统方法相比更加高效快捷,近年来不断被应用于制药、食品制品、环境工程等众多领域的菌种改良中。对genome shuffling近年来技术的优化及其应用进展进行综述。     

基于基因组改组技术选育多重抗性乙醇酵母

本文通过基因组改组（Genome shuffling）技术将酵母菌株的多重耐受性能集中于同一菌株,从而选育出多重抗性的乙醇酵母。以分别经过热灭活和紫外灭活的酵母GZ-0533、GZ-241、GZ一-582为亲本菌株,经过3轮基因改组后,筛选得到的28株酵母菌。第二次改组得到的MSR14和第三次改组得到的MSR23对多重胁迫条件的耐受性提高最多,乙醇产量分别达到8．58％和8．86％,较多重抗性最好的出发菌GZ-05相对提高了7％和10．5％。26SrDNA序列分析表明,MSR14和MSR23的26SrDNA序列与酿酒酵母（Saccharomyces cerevisiae）的同源性为100％,均认定为酿酒酵母。     

基因组重排技术及其在真菌育种中的应用

基因组重排(genome shuffling)是一种快速提高细胞表型的新颖的全基因组工程方法,它是基于多亲本的原生质体递归融合,提供了在缺少基因组序列数据或网络信息情况下的全基因组重组的优势。文中介绍了基因组重排的原理、优点及过程,概述了基因组重排在真菌育种中的应用,并展望了该技术的前景。     

无黑色素普鲁兰多糖出芽短梗霉的选育

为选育一株高产无黑色素普鲁兰多糖的出芽短梗霉(Aureobasidium pullulans),以A.pullulans As.40329和A.pullulans As.3#为出发菌株,通过紫外诱变(15 W)筛选出三株正突变菌株作为亲本。在此基础上,对三株亲本制备原生质体并采用35%的PEG4000介导进行全基因重组(Genome shuffling,GS)。随后,经双亲灭活筛选后获得A.pullulans的全基因重组菌F2-6。结果表明,菌株F2-6遗传性状稳定且发酵多糖产物中近乎无黑色素(OD654维持在0.02左右),多糖产量提高至(19.53±0.39)g/L,比原始菌As.3#提高119.69%,表明重组菌F2-6具有工业化生产普鲁兰多糖的潜能。     

微生物菌种选育中基因组重排技术应用的研究进展

基因组重排(genome shuffling)技术是通过传统诱变育种结合原生质体融合技术开发出的一种新型微生物菌种选育和改良技术,具有易操作、效率高和适用广等特点。本文对基因组重排技术的原理、操作过程及其在增加次生代谢产物产量、增强菌株耐受性和提高底物利用能力等方面的应用进行了综述。     

全新酵母基因组图谱出炉

来自美国斯坦福大学医学院遗传学系、斯坦福基因组技术研究中心（Stanford Genome Technology Center）、加拿大多伦多大学Terrenee Donnelly细胞与生物分子研究中心的研究人员设计出了一种新的方法对酵母中70000个核小体（Nucleosomes）成功进行了描绘，获得了一张全新的酵母基因组图谱，这有助于我们了解和预测细胞的核小体状态，以及研究细胞遗传物质活动过程中的详细细节。这一研究成果公布在《自然-遗传学》（Nature Genetics）杂志上。     

Genome Shuffling Enhanced e-poly-L-lysine Production of a Recombinant Streptomyces sp. Feel-1

利用Genomeshuffling技术对1株融合重组菌Streptomycessp．Feel-1产ε-聚赖氨酸（8-PL）进行了育种研究。首先对Feel-1进行了甲基磺酸乙酯（EMS）诱变,以甘氨酸（Gly）和S-2-氨乙基-L-半胱氨酸（AEC）作为抗性指标进行筛选,得到4株正向突变株,ε—PL摇瓶产量分别为1．78g／L、1．89g／L、1．85g／L、1．86g／L,比原始菌株Feel-1（1．60g／L）分别提高了11．3％、18．1％、15．6％、16．3％;然后将这4株产量提高株作为出发菌库,采用双亲灭活法进行了3轮递推式原生质体融合（Genome shuffling）,最终得到了3株产量大幅提高的重组菌,摇瓶产量分别为2．42g／L、2．35g／L和2．37g／L,比原始菌株分别提高了51．3％、46．9％和48．i％;选择其中1株（G3—67）进行补料分批发酵,192h时ε-PL产量达到32．2g/L。     

咖啡酰奎宁酸类化合物抑菌活性的网络药理学研究

目的:基于网络药理学方法探讨咖啡酰奎宁酸类化合物抑菌的作用机制。方法:通过文献挖掘和数据库检索获取咖啡酰奎宁酸类化合物对应靶点、抑菌相关靶点及两者交集靶点,将交集靶点构建蛋白互作（protein-protein interaction network,PPI）网络,并进行基因本体（Gene Ontology,GO）分析及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genome,KEGG）分析。结果:咖啡酰奎宁酸类化合物对应靶点483个,抑菌相关靶点805个,咖啡酰奎宁酸类化合物抑菌作用靶点75个,起关键作用的为TNF、AKT1、ALB、MMP9、EGFR、MAPK8等靶点。GO功能分析及KEGG通路富集分析结果显示,咖啡酰奎宁酸类化合物抑菌的作用机制主要涉及应激反应、代谢过程、生物调节、多细胞生物过程、细胞讯息传递等生物过程,并通过细胞外基质组织、胶原蛋白降解、基质金属蛋白酶的激活和神经生长因子信号通路等多条通路共同发挥作用。化合物与分子靶点对接结果良好,验证了网络构建预测的准确性。结论:本研究揭示了咖啡酰奎宁酸类化合物抑菌具有多靶点、多途径的特点,为其分子机制的进一步研究奠定了基础。     

Changes in smoking prevalence in Ukraine in 2001-5

Objectives

To analyse trends in smoking prevalence in Ukraine from three surveys conducted in 2001–5, and to explore correlates of observed changes, in order to estimate the stage of tobacco epidemic in Ukraine.

Design

Repeated national interview surveys in Ukraine in 2001, 2002 and 2005.

Main outcome measure

Prevalence of current smoking among the population aged ⩾15 years.

Results

The age‐standardised prevalence of current smoking in Ukrainian men was 54.8% in 2001 and 66.8% in 2005. Among Ukrainian women, prevalence increased from 11.5% in 2001 to 20.0% in 2005. ORs for yearly increase in prevalence were estimated as 1.164 (95% CI 1.111 to 1.220) for men and 1.187 (1.124 to 1.253) for women, which implies that, on average, 3–4% of men and 1.5–2% of women living in Ukraine join the smoking population each year.

Conclusions

In Ukraine, smoking prevalence is increasing in most population groups. Among men, the medium deprivation group with secondary education has the highest smoking prevalence. Among women, while the most educated, young and those living in larger cities are the leading group for tobacco use, other groups are also increasing their tobacco use. Tobacco promotion efforts appear to have been significantly more effective in Ukraine than smoking control efforts. The decrease in real cigarette prices in Ukraine in 2001–5 could be the main factor explaining the recent growth in smoking prevalence.Ukraine is a large eastern European country with high smoking prevalence. Tobacco products are widely available at very low prices, and the transnational tobacco industry is extremely successful in promoting its products and lobbying for its interests in the legislative field. Advocacy of tobacco control has only recently achieved some success in the legislative field, with the first tobacco control law being adopted in late 2005. Ukraine ratified the Framework Convention on Tobacco Control in 2006, but there is still much cause for concern regarding the enforcement and effectiveness of the adopted legislative measures. Unfortunately, the government is not yet involved in nationwide surveillance of the tobacco problem, and the available data have mostly been gathered with funding from foreign donors. Several attempts have been made to measure the extent of the tobacco epidemic in the countries of the former Soviet Union (FSU).^{1,2,3,4,5,6,7} Most of these studies have shown rather high smoking prevalence among men (50–70%) and comparatively low prevalence among women (5–20%). Most countries in the FSU have similar smoking rates, while certain trends are shown to be related to the differences in how the transnational tobacco industry succeeds in every national tobacco market.⁶ Unfortunately, few studies have been published showing trends in smoking prevalence in the FSU.⁷ In Ukraine, two studies^3,4 provided point estimates of smoking prevalence.The aim of this study was to analyse trends in smoking prevalence in Ukraine, on the basis of three surveys conducted in 2001–5, and to explore correlates of observed changes in order to estimate the stage of the tobacco epidemic in Ukraine.     

2010年河南省部分食品中重金属污染状况分析

了解河南省部分食品中重金属污染的现状。方法 在河南省18个省辖市采集8大类3 657份食品样品,按照国家标准检测食品中铅、镉和汞的含量,检测结果按照GB 2762—2012《食品安全国家标准食品中污染物限量》进行评价。结果 河南省部分食品中铅含量的平均值为0.34 mg/kg,总体超标率为8.8%(320/3 656),其中粮食、肉类和蔬菜中超标率较高,分别为17.2%(95/552)、14.5%(85/587)和10.7%(57/534);部分食品中镉含量的平均值为0.054 mg/kg,总体超标率为3.9%(143/3 657),其中食用菌和蛋类中超标率较高,分别为9.6%(49/508)和8.1%(42/518);部分食品中汞含量的平均值为0.077 mg/kg,总体超标率为8.3%(303/3 657),其中蔬菜和粮食中超标率较高,分别为25.3%(135/534)和12.3%(68/551)。结论 河南省部分食品中铅、汞污染情况较为严重,其中粮食、蔬菜和肉类食品中污染情况尤为突出,需要加强监管。     

Moulds and mycotoxins in sorghum stored in traditional containers in India

Various traditional containers have been used in India for storage of sorghum grains. Sorghum is susceptible to fungal infestation and toxin elaboration. The present study relates to the mould and mycotoxin contamination (aflatoxin B₁ and T-2 toxin) in stored sorghum in different storage containers viz. “Kotlu” (Storage rooms), earthenware pots, gunny bags and reed baskets. Aspergillus sp. and Fusarium sp. were the prominent genera and the “Kotlu” form of storage was most susceptible to fungal attack. Storage treatment had little effect on fungal contamination, but despite the fungal infestation, the mycotoxin contamination was found to be very low.     

中国固态发酵白酒中功能细菌研究进展

细菌是白酒酿造3大类微生物之一,其在白酒生产中有着重要的作用,如:芽孢杆菌、乳酸菌、放线菌和梭菌等,它们能产生酯类、有机酸、吡嗪、萜烯等微量成分,从而影响白酒的风味与品质。基于细菌在白酒中的重要功能以及微生物分离培养技术和分析检测技术的进步,近年来相关研究越来越深入。该文综述了近年来白酒功能细菌的研究进展,介绍了酿酒各个环节中主要功能细菌及其在白酒中的作用,旨在为白酒功能细菌的研究提供参考。     

2010年山西省食品中食源性致病菌监测分析

目的 了解山西省食源性致病菌的污染现状.方法 按照2010年度《全国食源性致病菌监测工作手册》进行.结果 1 576份样品中共检出阳性菌株149株,检出率为9.45％,其中单核细胞增生李斯特氏菌、副溶血性弧菌污染比较严重,总检出率为9.50％和8.22％,金黄色葡萄球菌、阪崎肠杆菌和沙门氏菌的总检出率分别为3.02％、1.11％和0.87％.不同类别食品中致病菌的检出率差异较大,生肉中致病菌的检出率高居榜首为49.33％,主要污染菌为沙门氏菌、单核细胞增生李斯特氏菌;沙拉中致病菌的检出率为17.39％,主要污染菌为单核细胞增生李斯特氏菌、金黄色葡萄球菌;生食水产品中致病菌的检出率为16.94％,主要污染菌为单核细胞增生李斯特氏菌、副溶血性孤菌;熟肉制品中致病菌的检出率为9.84％,主要污染菌为单核细胞增生李斯特氏菌、金黄色葡萄球菌、沙门氏菌;中式凉拌菜中致病菌的检出率为8.89％,主要污染菌为金黄色葡萄球菌;鲜榨果汁中致病菌的检出率为6.67％,主要污染菌为金黄色葡萄球菌;鲜冻水产品中致病菌的检出率为5.00％,主要污染菌为副溶血性弧菌;婴幼儿配方粉/米粉/谷粉/豆奶粉中阪崎杆菌的检出率为1.11％.结论 山西省多种食品均存在食源性致病菌不同程度的污染,应加大对散装食品、即食食品和生食水产品的监管.