米槠天然更新次生林皆伐地采伐剩余物叶分解及其化学组成变化

对三明35年生米槠天然更新次生林皆伐地采伐剩余物叶的分解速率以及化学组成的年变化进行研究.结果表明:2012年5月至2013年4月,叶分解速率随时间呈指数变化趋势,干质量损失率>80%.K发生净释放,残留率<5%;N先累积后释放,P呈现先释放后累积再释放的趋势,N和P的残留率分别为19%和16%.养分释放速率以K最大,其次为P,N最小.木质素浓度在2012年5—10月呈先降低后上升再降低的趋势,从2012年11月至试验结束无明显变化,而在整个分解过程中纤维素浓度变化不显著.采伐剩余物叶分解过程中,N/P先上升后下降再上升,比值由18.6上升到21.1,木质素/N在分解前期波动较大,后期变幅较小.     

云南黑老虎不同种源氨基酸和其他指标的分析与评价

为探讨黑老虎（Kadsura coccinea ）作为果用食物的营养价值,对采自云南省不同产地的野生黑老虎果实分别测定其果皮和果肉中的维生素 C、可溶性固形物、氨基酸、粗纤维、蛋白质、总糖等含量,并进行比较分析。结果表明：野生黑老虎果实含有丰富的人体必需的氨基酸,4个种源中,以勐海的黑老虎果肉中各人体必需氨基酸总含量最高,为0．256％,其次是屏边种源,为0．106％,景东种源为0．0943％,普文种源氨基酸总含量最少,为0．0274％。黑老虎果实还含丰富的维生素 C、粗纤维、蛋白质、总糖等。不同产地之间或单株之间各营养指标变化范围大,变异系数较高,优良种源或优良单株可选择性较强。果肉的养分灰关联度以勐海种源和第2单株最高,将作为备选优良种源和优良单株。     

嗜碱芽孢杆菌N16-5木寡糖结合蛋白XynE的功能表征

嗜碱芽孢杆菌(Bacillus sp.)N16-5是本实验室从内蒙古乌都淖湖沉积物中分离的嗜碱菌,含有丰富的多糖水解酶,能够利用广泛的单糖和多糖。实验室前期转录组研究发现其基因组上存在一个21 kb大小的木聚糖利用相关基因簇,其中包括xyn EFG基因簇编码的ABC转运蛋白。【目的】生物信息学分析预测xyn E编码转运蛋白的底物结合蛋白,通过敲除xyn E基因研究它对菌株N16-5利用木聚糖的影响。【方法】利用温敏型载体p NNB194介导的同源交换重组的方法构建了xyn E基因敲除菌株N16-5(Δxyn E),并通过基因回补对敲除菌株表型进行验证。通过检测菌株在木聚糖培养基中的生长情况及培养基中还原糖含量的变化来分析xyn E基因对菌株利用木聚糖的影响;通过HPLC检测分析不同培养时间点木聚糖培养基的组分,结合缺失菌株和野生型菌株在以木糖为唯一碳源的培养基中的生长情况来分析Xyn E所属ABC转运蛋白的底物特异性。【结果】相比野生型菌株,缺失型菌株N16-5(Δxyn E)在木聚糖培养基的生长曲线对数期明显延迟,最大生物量略低,且培养过程中出现了明显的还原糖的累积与消耗过程;回补菌株恢复了野生型表型,且最大生物量比野生型略高。HPLC检测分析显示,相比野生型菌株,缺失菌株培养过程底物消耗速度较慢,且16 h后出现明显的木四糖、木三糖和木二糖的累积,直至60 h后仍有较大量木二糖的存在;在木糖培养基中培养时,缺失型菌株和野生型菌株的生长趋势较一致。【结论】Xyn E蛋白特异性结合木寡糖,其所属ABC转运蛋白在嗜碱芽孢杆菌N16-5降解利用木聚糖过程中发挥着重要作用。     

美国鲥鱼对我国淡水生态系统的潜在入侵风险

近年来美国鲥鱼已被引入到我国的上海和广东等地进行水产养殖。本实验报道了在纯淡水人工养殖条件下美国鲥鱼的繁殖和生长情况。人工养殖鲥鱼的生长速度接近美国北卡罗莱纳州沿岸野生鲥鱼的生长速度,而高于特拉华河野生鲥鱼的生长速度;人工养殖的2龄鲥鱼既可以在养殖池中自然产卵,也可以通过注射激素人工诱导产卵,其平均受精率、孵化率和仔鱼17天内的存活率分别约15%、75%和54%。这一结果表明,尽管鲥鱼是在海水中进行繁殖的洄游性鱼类,但在生长环境适宜的淡水条件下同样可以正常生长,并达到性成熟和繁育后代,有可能在适宜的淡水水体中形成自我维持的种群。结合本实验结果和美国鲥鱼的其他生物学特性,本文认为美国鲥鱼在我国天然水体生态系统中具有潜在的入侵性。     

Insect Cell Culture and Biotechnology

The continued development of new cell culture technology is essential for the future growth and application of insect cell and baculovirus biotechnology. The use of cell lines for academic research and for commercial applications is currently dominated by two cell lines; the Spodoptera frugiperda line, SF21 (and its clonal isolate, SF9), and the Trichoplusia ni line, BTI 5B1-4, commercially known as High Five cells. The long perceived prediction that the immense potential application of the baculovirus-insect cell system, as a tool in cell and molecular biology, agriculture, and animal health, has been achieved. The versatility and recent applications of this popular expression system has been demonstrated by both academia and industry and it is clear that this cell-based system has been widely accepted for biotechnological applications. Numerous small to midsize startup biotechnology companies in North America and the Europe are currently using the baculovirus-insect cell technology to produce custom recombinant proteins for research and commercial applications. The recent breakthroughs using the baculovirus-insect cell-based system for the development of several commercial products that will impact animal and human health will further enhance interest in this technology by pharma. Clearly, future progress in novel cell and engineering advances will lead to fundamental scientific discoveries and serve to enhance the utility and applications of this baculovirus-insect cell system.     

Efficient Gene Transfer Mediated by HIV-1-based Defective Lentivector and Inhibition of HIV-1 Replication

Lentiviral vectors have drawn considerable attention recently and show great promise to become important delivery vehicles for future gene transfer manipulation. In the present study we have optimized a protocol for preparation of human immunodeficiency virus type-1 (HIV-1)-based defective lentiviral vectors (DLV) and characterized these vectors in terms of their transduction of different cells. Transient co-transfection of 293T packaging cells with DNA plasmids encoding lentiviral vector constituents resulted in production of high-titer DLV (0.5-1.2 × 107IU/mL), which can be further concentrated over 100-fold through a single step ultracentrifugation. These vectors were capable of transducing a variety of cells from both primate and non-primate sources and high transduction efficiency was achieved using concentrated vectors. Assessment of potential generation of RCV revealed no detection of infection by infectious particles in DLV-transduced CEM, SupT-1 and MT-2 cells. Long-term culture of transduced cells showed a stable expression of transgenes without apparent alteration in cellular morphology and growth kinetics. Vector mobilization to untransduced cells mediated by wild-type HIV-1 infection was confirmed in this test. Challenge of transduced human T-lymphocytes with wild-type HIV-1 showed these cells are totally resistant to the viral infection. Considering the effective gene transfer and stable gene expression, safety and anti-HIV activity, these DLV vectors warrant further exploration for their potential use as a gene transfer vehicle in the development of gene therapy protocols.     

扩散速率估算的影响

理论上,土壤呼吸通量的量值可以通过观测土壤呼吸CO₂扩散速率(&#601;c/&#601;t)计算得到。但是为获得&#601;c/&#601;t,通常须允许土壤呼吸箱内CO₂浓度升高,因此,如何估算外界大气CO₂浓度条件下的&#601;c/&#601;t是土壤呼吸观测技术的关键,关系到观测结果的准确性。通常&#601;c/&#601;t的估算会受土壤表层大气CO₂扩散梯度(即土壤呼吸箱内CO₂扩散梯度和大气CO₂浓度昼夜变化)的影响。目前,线性回归方法是土壤呼吸观测中估算&#601;c/&#601;t的基本方法。然而,常用的线性回 归方法会低估&#601;c/&#601;t,而指数回归方法则可以准确地估算&#601;c/&#601;t。夜间&#601;c/&#601;t的变化与大气CO₂ 浓度之间存在非常明显的负相关关系。夜间土壤表层大气CO₂扩散梯度的减小导致线性回归方法明显低估&#601;c/&#601;t。&#601;c/&#601;t的昼夜变化过程存在明显的非对称性现象,而指数回归方法可以更好地描述&#601;c/&#601;t昼夜变化的非对称性响应。     

PC-１基因表达增强C4-2B前列腺癌细胞生存

建立稳定表达外源PC-１基因的人前列腺癌骨转移C4-2B细胞模型,初步探讨PC- １基因表达对前列腺癌发展的影响.通过脂质体介导的方法,将融合PC-１基因的真核表达载体pcDNA3.1PC-１稳定转染C4-2B细胞,Western 印迹和RT-PCR技术,分别从蛋白水平和RNA水平确定外源PC-１基因表达. MTT和软琼脂集落形成能力等一系列方法,研究PC-１基因的功能,RT-PCR和实时定量PCR检测前列腺癌发生发展相关基因表达的变化. 结果表明,PC-１基因的高表达能够诱导雄激素受体（AR）调控基因和一系列重要的信号通路成员基因PSA、PSMA、NKX31、Jagged1、EphA3、SGEF和 NOTCH3等表达发生变化. 实验结果初步证明,PC-１基因表达在晚期前列腺癌中,以及在雄激素非依赖的转变中可以发挥作用,PC-１基因表达可调控一些重要信号通路.对PC-１基因功能深入研究将有可能为发现新的前列腺癌的诊断治疗分子靶标提供线索.     

RAD51 135G-->C modifies breast cancer risk among BRCA2 mutation carriers: results from a combined analysis of 19 studies

RAD51 is an important component of double-stranded DNA–repair mechanisms that interacts with both BRCA1 and BRCA2. A single-nucleotide polymorphism (SNP) in the 5′ untranslated region (UTR) of RAD51, 135G→C, has been suggested as a possible modifier of breast cancer risk in BRCA1 and BRCA2 mutation carriers. We pooled genotype data for 8,512 female mutation carriers from 19 studies for the RAD51 135G→C SNP. We found evidence of an increased breast cancer risk in CC homozygotes (hazard ratio [HR] 1.92 [95% confidence interval {CI} 1.25–2.94) but not in heterozygotes (HR 0.95 [95% CI 0.83–1.07]; P=.002, by heterogeneity test with 2 degrees of freedom [df]). When BRCA1 and BRCA2 mutation carriers were analyzed separately, the increased risk was statistically significant only among BRCA2 mutation carriers, in whom we observed HRs of 1.17 (95% CI 0.91–1.51) among heterozygotes and 3.18 (95% CI 1.39–7.27) among rare homozygotes (P=.0007, by heterogeneity test with 2 df). In addition, we determined that the 135G→C variant affects RAD51 splicing within the 5′ UTR. Thus, 135G→C may modify the risk of breast cancer in BRCA2 mutation carriers by altering the expression of RAD51. RAD51 is the first gene to be reliably identified as a modifier of risk among BRCA1/2 mutation carriers.     

贝莱斯芽孢杆菌抗真菌能力及基因组分析北大核心CSCD

蛋白水解酶及脂肽类化合物具有广谱的抑菌活性,这些抑菌活性蛋白及物质拥有巨大的研究和开发潜力。本研究通过蛋白水解酶平板筛选到一株对6种不同病原真菌有较明显抑菌效果的菌株,命名为X49,对该菌株进行理化分析,并由16S rRNA基因测序、API测试和电镜分析鉴定菌种,进一步分析其基因组序列,再利用偶氮酪蛋白方法分析蛋白水解酶活性及提取脂肽类化合物进行抗真菌测试。结果表明,该菌株在10–50℃、pH 4.0–9.0范围内均可以生长,对环境盐浓度的适应性较强,在10%NaCl浓度下仍然能够生长。菌种鉴定发现X49菌株属于贝莱斯芽孢杆菌(Bacillus velezensis)。基因组分析则显示,B. velezensis X49菌株有11个编码的丝氨酸蛋白酶,其基因编号1_894属于S8枯草杆菌蛋白酶家族,与已知具有抗真菌能力的丝氨酸蛋白酶有高达99%的相似度。另外有多达30个编码的非核糖体肽合成酶,其中包含参与合成已知的表面活性素、伊枯草菌素、丰原素、杆菌肽及短杆菌肽脂肽类化合物。胞外蛋白水解酶活性分析发现其在高温下仍具有一定的活性。将B. velezensis X49菌株与中药干姜共发酵后,提取出的脂肽类化合物,其抑菌功效大大提高。由此可见,本研究筛选出的B. velezensis X49菌株对于植物及人类病原菌都有很显著的抑制作用。而与中药共发酵的活性物质可作为未来药物的研究开发。