黄河下游平原不同非农生境中植物多样性

自然、半自然等非农生境是区域农业景观的重要组成部分,对区域生物多样性保护具有重要意义。黄河下游平原区是典型的农业景观,农田是最主要的景观类型,林地、树篱等景观要素散布其中,为研究区内非农生境中生物多样性及其生态效应,采用典型样地法对区内典型农业景观中林地、树篱、田间道路和沟渠等主要非农生境中的植物群落进行调查研究。结果显示:区内植物组成以菊科、禾本科等为主;区内植物科的地理成分以世界分布和泛热带分布为主,各生境间有一定的差异,属的地理成分复杂,具有中国15个种子植物属分布型中的13个,以温带分布、世界分布和泛热带分布为主,总体上区内的植物组成以广布种为主,优势科属明显,特有种缺乏;各非农生境中的植物多样性存在一定的差异,林地和树篱生境具有较高的物种丰富度和多样性,显著高于田间道路,沟渠、林地和树篱生境中的物种均匀度和群落盖度均显著高于田间道路;β多样性分析表明田间道路生境中的群落组成分化程度在各样点间最大(β多样性指数最高);树篱、林地和田间道路等生境间群落相似性均较高,但其群落结构和优势种组成方面却存在显著的差异,沟渠作为一种特殊生境与其它生境间的群落相似性相对较低。研究表明,在黄河下游平原典型农业景观中,作为非农生境存在的林地和树篱在物种多样性维持中具有重要地位,沟渠为水生和湿生植物提供了庇护所,意义重大;各生境间高的群落相似性仅是物种组成名录相似性的反映,其空间格局和优势种群间差异明显,各生境植物群落的生态功能差异巨大。未来区内生物多样性的保护应重在生态系统过程、功能的加强以及生态系统服务的维持和提高,且需进一步在景观水平上探讨各非农景观要素的空间构型对其生态效益的影响机制及其调控和管理策略。     

牛β-乳球蛋白基因的克隆及部分序列测定

为了制备乳腺生物反应器所需要的乳腺特异性表达的调控序列,用ＰＣＲ法从奶牛染色体上分５段扩增出了全长的牛 ＢＬＧ基因约 ８ ｋｂ,包括 １．８ ｋｂ的 ５’侧翼区、１．７ ｋｂ的 ３’侧翼区及 ４．７ ｋｂ的ｇＤＮＡ区。扩增出的各片段克隆到Ｔ－Ｖｅｃｔｏｒ上,酶切鉴定及序列分析均证实了所扩增片段的正确性。     

农业景观中非农景观要素结构特征对植物物种多样性的影响——以封丘县为例

农业景观中的非农生境对维持与提高农业景观的生物多样性具有非常关键的作用。为了探究非农生境的相关结构属性对农业景观中植物物种多样性的影响,选择黄河下游平原区的封丘县为研究区域,对研究区内42个样点的非农生境进行植物多样性调查,并对各个样点周围1 km范围内的非农景观要素进行了提取,分析不同非农生境中植物物种组成及其景观要素的构成、结构及空间配置对植物物种多样性的影响。研究结果表明:不同类型的非农生境中,物种组成共有种相对较多,特有种或指示种较少;林地与树篱具有相对较高的物种多样性,以沟渠为生境的植物物种组成与其它两种生境类型相比存在明显差异;林地与树篱/沟渠的组成比例相当时,植物物种丰富度最高;景观指数对不同非农生境中的植物物种具有明显影响,景观破碎化及人为干扰指数的影响较为显著。未来在对本区域内农业景观进行结构优化的过程中,应从非农景观要素的改造入手。通过调整和设置非农景观要素的不同类型及比例、合理改造其结构与空间配置,为最终实现农业景观的有效管理与可持续健康发展奠定重要的研究基础。     

微生物功能基因组学研究

自从1995年流感嗜血杆菌的基因组序列测定完成之后[1],目前已有75种(株)微生物的基因组完成测序,160多种(株)微生物的基因组测序正在进行中[2]。随着各种微生物基因组测序工作的不断完成和序列信息的积累,微生物基因组学研究的重点已由结构基因组学向功能基因组学转移。微生物功能基因组学研究不仅要阐明微生物基因组内每个基因的作用或功能,还要研究基因的调节及表达谱,进而从整个基因组及其全套蛋白质产物的结构、功能、机理的高度去了解微生物生命活动的全貌,揭示微生物世界的各种前所未知的规律,并使之为人类和社会服务。与真核生物相比,虽然微生物的基因组相对简单,但微生物基因组学研究仍具有重大的科学和经济意义。在细菌基因组中,既有编码在极端环境下起催化作用的酶的基因,也有编码分解化学污染物的酶的基因,这些基因在真核细胞是不存在的。通过微生物功能基因组学研究,还能发现药物靶位和疫苗抗原。微生物基因的功能及表达研究结果也能为研究复杂生物的基因功能提供参考。近些年微生物功能基因组学研究受到了普遍重视。日本组织了十几所大学和研究机构,计划用5年时间完成大肠杆菌的功能基因组研究[3]。日本还与欧洲联合正在开展枯草杆菌功能基因组学研究[4]。其它微生物的功能基因组学研究也在进行中。由于微生物的种类繁多,功能基因组研究的内容又较丰富,要全面介绍微生物功能基因组学研究是困难的。本文仅从未知功能基因的鉴定、药物靶位及疫苗抗原研究、致病机制研究、生物功能图谱研究4个方面进行简要的评述。     

重组人GM－CSF／MCAF融合蛋白抗血清的制备与鉴定

重组人粒细胞巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）和人单核细胞趋化激活因子（ＭＣＡＦ）融合蛋白经ＳｅｐｈａｄｅｘＧ－７５和ＣＭ－ＳｅｐｈａｒｏｓｅＦＦ两步柱层析,获得了电泳纯的ＧＭ－ＣＳＦ／ＭＣＡＦ融合蛋白。为进一步研究其结构与功能,我们以纯化的该融合蛋白为抗原免疫家兔制备抗血清。ＤｏｔＥＬＩＳＡ和Ｗｅｓｔｅｒｎｂｌｏｔ试验表明,该抗血清效价高、特异性好,可分别与ＧＭ－ＣＳＦ／ＭＣＡＦ、ＧＭ－ＣＳＦ和ＭＣＡＦ发生反应。     

Construction of a trivalent candidate Shigella vaccine strain with host-vector balanced-lethal system

A trivalent liveShigella vaccine candidate FSD01 against S.flexneri 2a, S.sonnei and S.dysenteriae I was constructed. This candidate strain was based on the S.flexneri 2a vaccine T32. By homologous recombination exchange, the chromosomalasd gene of T32 was site-specifically inactivated, resulting in the strain unable to grow normally in LB broth, while anotherasd gene of S.mutans was employed to construct an Asd⁺ complementary vector. This combination ofasd ^- host/Asd⁺ vector formed a balanced-lethal expression system in T32 strain. By use of this system, two important protective antigen genes coding for S.sonnei Form I antigen and Shiga toxin B subunit were cloned and expressed in T32, which led to the construction of trivalent candidate vaccine FSD01. Experimental results showed that this strain was genetically stable, but its recombinant plasmid was non-resistant. Moreover, it was able to effectively express trivalent antigens in one host and induce protective responses in mice against the challenges of the above threeShigella strains.     

水稻糯性相关基因的全基因组关联分析

利用1 090 071个SNP标记, 对419份广西地方稻种资源核心种质的糯性进行全基因组关联分析。结果表明, 运用混合线性模型, 在P<4.72×10^-8 (4.72E-8)水平下, 检测到45个与糯性显著关联的SNP位点, 均位于第6号染色体上。通过筛选显著关联位点上、下游各150 kb区域内的基因, 共找到305个候选基因, 其中包含2个与淀粉合成酶相关的Wx和SSIIa基因; 同时在第6号染色体5.69-5.89 Mb区域发现4个与糯性显著关联的SNP位点, 该区域可能是影响水稻(Oryza sativa)糯性的重要候选区域。     

Overexpression of Ras Homologous C (RhoC) Induces Malignant Transformation of Hepatocytes In Vitro and in Nude Mouse Xenografts

Ras homologous C (RhoC) is expressed in various cancers, including hepatocellular carcinoma (HCC). In this study, we first analyzed RhoC expression in 46 HCC tissue specimens and found that RhoC expression was significantly increased in HCC tissues compared to the adjacent normal liver tissues. Next, we investigated the role of RhoC in malignant transformation of normal hepatocytes. The HL7702 cell line was stably transfected with a RhoC expression vector and then subjected to cell proliferation, differentiation, colony formation, migration and invasion assays, as well as nude mouse xenograft assays. Gene expressions in these cells were determined using RT-PCR and Western blot. Overexpression of RhoC significantly promoted proliferation and anchorage-independent growth of HL7702 cells, but suppressed cell differentiation, as compared with the parental cells and the empty vector-transfected control cells. Moreover, RhoC overexpression induced migration and invasion of HL7702 cells in vitro. Molecularly, RhoC increased the expression of cell cycle-related genes, matrix metalloprotease 2 (MMP2), MMP9 and vascular endothelial growth factor (VEGF). In addition, RhoC-transfected cells formed tumors in nude mice, whereas vector-transfected HL7702 cells did not form any tumors in nude mice. This study demonstrated the role of RhoC overexpression in malignant transformation of normal human hepatocytes, suggesting that RhoC may function as an oncogene in hepatocytes.     

Construction of a recombinant human GM-CSF/MCAF fusion protein and study on its in vitro and in vivo antitumor effects

A novel cytokine fusion protein was constructed by fusing granulocyte macrophage colony stimulating factor (GM-CSF) with monocyte chemotactic activating factor (MCAF), which acts as a factor directing effector cells (monocytes) to a target site. The recombinant human GM-CSF/MCAF fusion protein could sustain the growth of GMCSF-dependent cell line TF1 and was chemotactic for monocytes. Thein vitro antitumor effect showed that rhGM-CSF/MCAF could activate monocytes to inhibit the growth of several human tumor cell lines, including a promyelocyte leukemia cell line HL-60, a lung adenocarcinoma cell line A549, a hepatoma cell line SMMC-7721 and a melanoma cell line Bowes. Furthermore, the cytotoxicity of monocytes activated by rhGM-CSF/MCAF against HL-60 and A549 was greater than that activated by GM-CSF or MCAF alone, even greater than that activated by a combination of GM-CSF and MCAF, suggesting that the fusion protein has synergistic or enhanced effects. Thein vivo antitumor effect indicated that rhGM-CSF/MCAF had marked antitumor effect against A549 tumor in nude mice and even completely suppressed tumor formation. rhGM-CSF/MCAF was significantly more effective in inhibiting tumor growth than rhGM-CSF. Histological analysis showed that tumor site injected with rhGM-CSF/MCAF was infiltrated by a large number of monocytes while a sparse infiltration of monocytes was observed at the tumor site injected with rhGM-CSF or normal saline, suggesting that the antitumor effect of rhGM-CSF/MCAF was mediated by the recruitment of a large number of monocytes to the tumor site.