Application of radiation hybrid in gene mapping

Ｒａｄｉａｔｉｏｎｈｙｂｒｉｄ(ＲＨ)ｍａｐｐｉｎｇｉｓａｓｏｍａｔｉｃｃｅｌｌｇｅｎｅｔｉｃｍａｐｐｉｎｇｔｅｃｈｎｉｑｕｅｗｉｔｈａｒｅｓｏｌｕｔｉｏｎｏｆａｂｏｕｔ500ｋｂ.Ｉｔｈａｓｂｅｃｏｍｅａｇｅｎｅｒａｌｗａｙｔｏｃｏｎｓｔｒｕｃｔｈｉｇｈｒｅｓｏｌｕｔｉｏｎ,ｃｏｎｔｉｇｕｏｕｓｐｈｙｓｉｃａｌｍａｐｏｆｈｕｍａｎｃｈｒｏｍｏｓｏｍｅｓ[1].ＢａｓｅｄｏｎｅａｒｌｉｅｒｓｔｕｄｉｅｓｏｆＧｏｓｓａｎｄＨａｒｒｉｓ[2]ａｎｄｍｏｄｉｆｉｃａｔｉｏｎｌａｔｅｒｂｙＣｏｘａｎｄｃｏｗｏｒｋｅｒ…     

瘤胃降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物的分离鉴定

【目的】本试验从瘤胃中分离鉴定降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物,为深入探究甲烷菌对厌氧真菌代谢途径的影响及相关调节机制奠定基础。【方法】利用厌氧滚管技术从荷斯坦奶牛瘤胃内容物中分离厌氧真菌与甲烷菌共培养物,通过形态学观察和DAPI染色以及甲烷菌16S rRNA基因序列分析方法分别对厌氧真菌及甲烷菌进行鉴定。【结果】从荷斯坦奶牛瘤胃中共分离到28株厌氧真菌与甲烷菌共培养物。共培养物中的厌氧真菌均为单中心菌株,分别属于Piromyces,Neocallimastix和Caeomyces属,所占百分比为53.57%,42.86%及3.57%。甲烷菌16S rRNA基因序列分析结果表明,共培养物中的甲烷菌均为甲烷短杆菌。本研究共获得四种不同的厌氧真菌与甲烷菌组合,分别为Piromyces/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter olleyae菌株,Neocallimastix/类Methanobrevibacter thaueri菌株及Caecomyces/类Methanobrevibacter olleyae菌株,分别占总数的53.57%,39.29%,3.57%及3.57%。【结论】分离得到的28株厌氧真菌和甲烷菌共培养物中,占优势的为具有丰富丝状假根的厌氧真菌Piromyces和Neocallimastix以及类Methanobrevibacter olleyae属的甲烷短杆菌。本研究为进一步研究瘤胃内厌氧真菌与甲烷菌相互代谢关系奠定基础。     

杂交水稻及其“三系”线粒体DNA的AP—PCR指纹图谱

为了研究水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）细胞质雄性不育（ＣＭＳ）与线粒体基因组的关系,应用ＡＰ－ＰＣＲ 分析,用７ 个任意单引物对６ 种水稻品系线粒体ＤＮＡ 进行了扩增。水稻线粒体ＤＮＡ 的ＡＰ－ＰＣＲ 产物可分为三种类型：（１）所有供试品系均能扩增的片段,它们代表了线粒体ＤＮＡ 在进化上的保守性序列。有４ 个引物检测到这类片段。（２）２ 个以上水稻品系共同出现而在全部供试材料间存在差异的扩增片段,这类片段是检测水稻线粒体ＤＮＡ多态性的主要来源。（３）一种细胞质类型所特有的扩增片段,从引物Ｒ２ 和Ｖ５ 的扩增产物中发现了这类片段,它们可能与ＣＭＳ有关联。另外,ＷＡ型不育系珍汕９７Ａ 与其杂种之间在６ 个引物的扩增图谱上均存在不同程度的差异,说明两者的线粒体ＤＮＡ序列结构可能存在某种差别     

放牧对小叶锦鸡儿种子产量的影响

锡林河流域草地灌木化表现为小叶锦鸡儿（Caragana microphylla）种群的扩张。为了研究放牧与小叶锦鸡儿种群扩张的关系, 从小叶锦鸡儿的种子产量和萌发入手, 分析放牧对小叶锦鸡儿生殖的直接影响（即放牧的直接影响）以及不同放牧处理下昆虫种群对小叶锦鸡儿生殖造成的影响（即放牧的间接影响）。通过比较不同处理下小叶锦鸡儿果荚的数量、种子产量以及昆虫对种子的取食, 发现放牧产生的直接影响和间接影响性质不同, 放牧的直接影响是不利于小叶锦鸡儿的繁殖,而其间接影响却对小叶锦鸡儿的繁殖有利, 且这两种影响在很大程度上会互相抵消。根据所得的研究结果可知在停止放牧之后的一段时间内, 小叶锦鸡儿种群扩张的可能性会增加, 因此在草地生态系统管理中应该注意这个问题。     

变性梯度凝胶电泳法研究断奶仔猪粪样细菌区系变化

利用PCR和DGGE技术分析了12头仔猪在断奶后其粪样细菌区系的变化。粪样细菌16S rDNA的V6～V8可变区经PCR扩增,扩增产物经DGGE电泳后再进行相似性分析。结果表明,仔猪断奶当天粪样 DGGE谱带少,同窝仔猪间图谱相似。断奶后,随着断奶时间的推移,每头仔猪的DGGE图谱带逐渐增多,变得复杂和多样,仔猪个体间DGGE图谱差异逐渐增大。仔猪是否同窝以及所采食日粮类型对DGGE图谱没有明显影响。相似性分析还表明,日粮中添加寡果糖的仔猪在断奶后第1周,其粪样微生物区系变化迅速,而后缓慢。     

Surface Display of Human Serum Albumin on Bacillus subtilis Spores for Oral Administration

Human serum albumin (HSA) is the major protein component of human plasma. To date, HSA for clinical uses is mostly produced by fractionation of human whole blood, which is accompanied by a lot of limitations. To obtain long-term bioactive albumin, we used hsa as a foreign gene and constructed a recombinant plasmid pJS700-HSA which carries a recombinant gene cotC-hsa under the control of cotC promoter. Plasmid pJS700-HSA was transformed into Bacillus subtilis by double cross-over and an amylase inactivated mutant was produced. After induction of spore formation, western blot and fluorescence immunoassay were used to monitor HSA surface expression on spores. We estimated that HSA displayed on the spore accounted for 0.135 % of the total spore proteins and about 0.023 fg HSA were exposed on the surface of each spore. Oral administration to mice with spores displaying HSA implied that the recombinant spores may have potential ability to increase the serum albumin level in vivo due to the resistant characters of spores.     

hnRNP C and polypyrimidine tract-binding protein specifically interact with the pyrimidine-rich region within the 3'NTR of the HCV RNA genome

Like other members of the Flaviviridae family, the 3' non-translated region (NTR) of the hepatitis C virus (HCV) is believed to function in the initiation and regulation of viral RNA replication by interacting with components of the viral replicase complex. To inves-tigate the possibility that host components may also participate in this process, we used UV cross-linking assays to determine if any cellular proteins could bind specifically to the 3'NTR RNA. We demonstrate the specific interaction of two host proteins with the extensive pyrimidine-rich region within the HCV 3'NTR. One host protein migrates as a doublet with a molecular weight of 57 kDa and is immunoreactive with antisera specific for polypyrimidine tract-binding protein (PTB), and the other protein (35 kDa) is recognized by a monoclonal antibody specific for heterogeneous nuclear ribonucleoprotein C (hnRNP C). These results suggest that recognition of the large pyrimidine-rich region by PTB and hnRNP C may play a role in the initiation and/or regulation of HCV RNA replication.