PCR-DGGE的原理及在动物肠道菌群分析中的应用

肠道菌群是机体常在菌群的重要组成部分,机体的健康和疾病的发生和发展与其多样性及种群的变化有着密切关系。因此,对肠道菌群进行研究具有重要意义。变性梯度凝胶电泳(DGGE)主要是基于突变型和野生型核酸序列的不同而导致其变性浓度的差异,利用变性梯度胶进行分离。它是一种分析微生物群落的有效工具,可以用于研究肠道菌群结构多样性和种群结构的变化。作者对PCR-DGGE的原理、优缺点及在动物肠道菌群分析中的应用和前景作一简要综述。     

DGGE技术在瘤胃微生物多态性研究中的应用

变性梯度凝胶电泳(DGGE)技术是一种新型的用于检测核酸变异和点突变的电泳方法,具有可靠性强、重现性高、方便快捷等优点,可应用于瘤胃微生物群落多样性和动态性分析.本文对DGGE技术的原理、优点、在瘤胃微生物群落多样性和动态性分析方面的应用结果及今后的发展方向进行了全面论述,为更好地认识和应用该技术提供了参考.     

DGGE技术在瘤胃微生物多态性研究中的应用

利用传统的方法很难鉴别瘤胃微生物的差异,变性梯度凝胶电泳（DGGE）技术就很好地解决了该问题。DGGE是一种新型的用于检测核酸变异和点突变的电泳方法,具有可靠性强、重现性高、方便快捷等优点,可应用于瘤胃微生物群落多样性和动态性分析。     

DGGE技术在瘤胃微生物多态性研究中的应用

利用传统的方法很难鉴别瘤胃微生物的差异,变性梯度凝胶电泳(DGGE)技术就很好地解决了该问题.DGGE是一种新型的用于检测核酸变异和点突变的电泳方法,具有可靠性强、重现性高、方便快捷等优点,可应用于瘤胃微生物群落多样性和动态性分析.     

苏云金芽胞杆菌在小鼠肠道的消长及对小鼠肠道菌群的影响

利用选择性培养平板计数法,研究了苏云金芽胞杆菌(Bt)营养体和芽胞在小鼠肠道的数量消长及对小鼠肠道生理性有益菌群(大肠杆菌、双歧杆菌和乳酸杆菌)数量的影响,同时利用基于16S rDNA作为分子标记的变性梯度凝胶电泳(DGGE)研究了Bt对小鼠肠道微生物多样性的影响。结果表明:Bt营养体和芽胞在小鼠肠道内很快死亡,也未对小鼠肠道有益菌群数量和细菌多样性产生不良影响。说明苏云金芽胞杆菌(Bt)营养体及芽胞对小鼠是安全的。     

DGGE技术的原理及其在动物胃肠道微生态系统研究中的应用

利用传统的培养方法很难将动物胃肠道的所有菌群进行培养,因此也很难鉴别各个时期动物胃肠道微生物之间的差异。变性梯度凝胶电泳(DGGE)技术可以避免传统方法的弊端,进而可以进一步确定动物胃肠道微生态系统的差别以及菌群的多样性。主要综述了DGGE技术的原理﹑优缺点以及在动物胃肠道微生态系统研究中的应用和前景。     

变性梯度凝胶电泳及其在微生态领域应用的研究进展

综述了不依赖于培养的分子生物学方法变性梯度凝胶电泳的原理、步骤、优点以及变性梯度凝胶电脉的不是及解决方法。对变性梯度凝胶电脉在环境微生物,动物肠道的多样性及微生物菌群动态变化及前景进行了研究和阐述,并指出了变性梯度凝胶电泳具有良好的发展前景。     

变性梯度凝胶电泳及其在微生态领域应用的研究进展

综述了不依赖于培养的分子生物学方法变性梯度凝胶电泳的原理、步骤、优点以及变性梯度凝胶电脉的不是及解决方法.对变性梯度凝胶电脉在环境微生物,动物肠道的多样性及微生物菌群动态变化及前景进行了研究和阐述,并指出了变性梯度凝胶电泳具有良好的发展前景.     

DGGE技市在动物胃肠道微生态研究中的应用

变性梯度凝胶电(Denaturing Gradient Gel Electrophoresis,DGGE)术是通过直接分离核酸片段对微生物群落进行研究,由于其具有可靠性强、重复性好、方便快捷等优点.被广泛应用于微生物群落多样性和动态性分析,并成为微生物分子生态学研究中的热点技术之一.本文主要介绍DGGE技术的原理、工作流程及优缺点.并着重分析了其在动物胃肠道微生态研究中的应用.     

DGGE技术及其在瘤胃微生物多样性研究中的应用

基于可靠性强、重现性高、方便快捷等优点,变性梯度凝胶电泳（DGGE）技术已经成为微生物群落多样性和动态分析的强有力工具。本文在阐述PCR—DGGE原理及流程的的基础上,对DGGE技术在瘤胃微生物多样性研究中的具体应用及其局限性与改进方法进行了全面阐述。     

Determination of Ileum Microbial Diversity of Broilers Fed Triticale- or Corn-Based Diets and Colonized by Salmonella

Diversity of the bacterial communities in the ileum of broilers was characterized using denaturing gradient gel electrophoresis. Denaturing gradient gel electrophoresis separation of polymerase chain reaction amplicons of the V2–V3 variable regions of the 16S rDNA is a common method to profile community diversity and has been used to assess the effects of diet and antibiotics on the ileal bacterial community of chickens. Broilers raised either on litter floor or in cage batteries were fed either a finely ground corn- (control), a finely ground triticale-, or a whole triticale-based diet from 0 to 42 d. Microbial DNA was extracted from the ileum content of 42-d-old broilers, and the 16S rDNA gene was amplified by polymerase chain reaction and the amplicons separated by denaturing gradient gel electrophoresis. Diversity indexes including richness, evenness, diversity, and pairwise similarity coefficients were calculated. Diversity indexes were related to the dietary treatments, housing designs, and to changes in Salmonella colonization of broiler ceca as characterized by the most probable number method. Higher microbial diversity indexes were observed among birds fed whole triticale-based diets and reared on litter floors. In contrast, finely ground grain treatments had lower diversity and higher Salmonella prevalence than the whole triticale treatment. The data indicated that combination of high dietary fiber content and increased coarseness of the diet by feeding whole triticale stimulated microbial community diversity and discouraged Salmonella colonization, perhaps through a competitive exclusion-type mechanism.     

分子生物技术在牛奶微生物群落研究中的应用

目前,奶牛乳房炎病原学研究的主要方法为对牛奶中的微生物进行传统的分离鉴定。由于这些微生物的形态过于简单,可提供的信息少,而且很多无法进行纯培养,传统方法已无法满足对其群落进行全面的客观认识。随着现代分子生物学技术的发展,随机扩增多态性DNA标记、变性梯度凝胶电泳、单链构象多态性检测、实时荧光定量PCR、核酸探针等技术为牛奶中微生物群落的研究提供了更为有效、直观的方法,更进一步揭示了牛奶中微生物的种群结构。     

Manipulation of the gut microbiota in C57BL/6 mice changes glucose tolerance without affecting weight development and gut mucosal immunity

Inflammatory diseases such as type 2 diabetes (T2D) in humans and mice are under the influence of the composition of the gut microbiota (GM). It was previously demonstrated that treating Lep(ob) mice with antibiotics improved glucose tolerance. However, wild type C57BL/6J mice may also exhibit plasma glucose intolerance reminiscent of human T2D. We hypothesized that antibiotic treatment in C57BL/6 mice would have an impact on glucose tolerance without affecting weight and gut immunology. When compared to mice treated with erythromycin or the controls, treatment for five weeks with ampicillin improved glucose tolerance without significantly affecting the weight or the number of gut mucosal regulatory T cells, tolerogenic dendritic cells or T helper cells type 1. 16S rRNA gene based denaturing gradient gel electrophoresis profiles clearly clustered according to treatment and showed that antibiotic treatment reduced GM diversity. It is concluded that antibiotic treatment changes glucose metabolism as well as the composition of the GM in C57BL/6 mice, and that this does not seem to be correlated to weight development in the mice.     

Fermentation and microbial population dynamics during the ensiling of native grass and subsequent exposure to air

To study the microbial population and fermentation dynamics of large needlegrass (LN) and Chinese leymus (CL) during ensiling and subsequent exposure to air, silages were sampled and analyzed using culture‐based techniques and denaturing gradient gel electrophoresis (DGGE). A total of 112 lactic acid bacteria (LAB) strains were isolated and identified using the 16S rRNA sequencing method. Lactic acid was not detected in the first 20 days in LN silage and the pH decreased to 6.13 after 45 days of ensiling. The temperature of the LN silage increased after approximately 30 h of air exposure and the CL silage showed a slight temperature variation. Enterococcus spp. were mainly present in LN silage. The proportion of Lactobacillus brevis in CL silage increased after exposure to air. LN silage with a higher proportion of Enterococcus spp. and propionic acid concentration did not show higher fermentation quality or aerobic stability than CL silage, which had a higher concentration of acetic acid, butyric acid and increased proportion of L. brevis after exposure to air.     

家鹑与野生日本鸣鹑群体微卫星DNA标记的遗传学分析

选择10个微卫星标记位点，合成引物对微山湖野生日本鸣鹑和家鹌鹑各40只进行PCR扩增，变性聚丙烯酰胺凝胶电泳检测分型，并对其品种内和品种间的遗传变异进行群体遗传学分析，从分子水平上揭示这两个鹌鹁群体的遗传结构关系、亲缘关系和遗传分化程度，为鹌鹑的遗传资源研究提供新资料。     

日粮中添加茶皂素和豆油对羔羊瘤胃细菌区系的影响

研究茶皂素和豆油对羔羊瘤胃细菌区系的影响。选16只健康断奶湖羊,随机分入对照、茶皂素(TS)、豆油(SO)、茶皂素和豆油混合添加组(TS-SO)。饲喂60 d后屠宰取瘤胃液,进行变性梯度凝胶电泳(DGGE)分析。结果表明:日粮中添加TS和SO对瘤胃细菌区系有一定的影响,TS-SO组细菌多样性增加,但各组间DGGE条带数差异不显著(P0.05)。切胶、测序分析图谱中4个特异条带分属于苍白杆菌、不动菌、葡萄球菌和甲基杆菌属,相似性均达99%以上。表明日粮中添加茶皂素和豆油可以一定程度改变羔羊瘤胃细菌区系,但对细菌多样性影响不显著。     

应用PCR-DGGE分析鸭场土壤细菌群落结构

为了解贵州省三穗县8个养鸭场环境土壤中细菌群落结构多样性概况,本试验采用试剂盒提取土壤样品细菌总DNA,PCR扩增细菌16S rDNA V3可变区,并通过变性梯度凝胶电泳(DGGE)技术对扩增产物进行凝胶分离,回收测序共得到43条特异性条带,其中7、8号养鸭场条带数相对较少.DGGE图谱经Quantity One软件分析,结果显示,同一鸭场样品相似性存在差异,绝大部分鸭场样品间相似性均高于50%,所测序列与GenBank中相应原核生物16S rDNA序列经同源性比对可见相似性在90%~100%之间,共包括5个大纲的细菌种类:变形菌门(Proteobacteria)的α,β类群、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)和厚壁菌门(Firmicutes).