An extracellular poly (3-hydroxybutyrate) depolymerase from Penicillium sp. DS9713a-01

Summary Penicillium sp. DS9713a-01 was obtained by ultraviolet (u.v.) light mutagenesis from the Penicillium sp. DS9713a which can degrade poly (3-hydroxybutyrate) (PHB). The enzymatic activity of DS9713a-01 was 97% higher than that of the wild-type strain. The DS9713a-01 mutant could completely degrade PHB films in 5 days; however, the wild-type strain achieved only 61% at the same time. The extracellular PHB depolymerase was purified from the culture medium containing PHB as the sole carbon source by filtration, ammonium sulfate precipitation and chromatography on Sepharose CL-6B. The molecular weight of the PHB depolymerase was about 15.1kDa determined by SDS-polyacrylamide gel electrophoresis. The optimum activity of the PHB depolymerase was observed at pH 8.6 and 50 °C. The enzyme was stable at temperatures below 37 °C and in the pH range from 8.0 to 9.2. The activity of PHB depolymerase could be activated or inhibited by some metal ions. The apparent K _m value was 0.164 mg ml⁻¹. Mass spectrometric analysis of the water-soluble products after enzymatic degradation revealed that the primary product was the monomer, 3-hydroxybutyric acid.     

樟科濒危植物思茅木姜子遗传多样性的ISSR分析

本文采用ISSR标记对中国特有且仅在云南南部狭域分布的樟科濒危植物思茅木姜子(Litseaszemaois)现存8个居群的遗传多样性进行了研究。从96条引物中筛选出了10条,对103个个体进行了扩增,共扩增出77条条带,其中多态性条带为67条。分析结果表明:(1)思茅木姜子的遗传多样性水平很高。在物种水平上,多态位点百分率PPB=87.01%,平均每个位点的有效等位基因数Ne=1.4006,Nei’s基因多样度指数H=0.2466,Shannon多样性信息指数Hsp=0.3826;在居群水平上,PPB=37.99%,Ne=1.2500,H=0.1418,Shannon多样性信息指数Hpop=0.2088。(2)居群间的遗传分化较低。基于Nei’s遗传多样性分析得出的居群间遗传分化系数Gst=0.3700;Shannon’s居群分化系数((Hsp–Hpop)/Hsp)为0.45。AMOVA分析显示:思茅木姜子的遗传变异主要存在于居群内,占总变异的72.99%,居群间的遗传变异占27.01%,表明思茅木姜子属于异交种。(3)两两居群间的Nei’s遗传一致度(I)的范围为0.8233–0.9761。经Mantel检测,居群间的遗传距离和地理距离之间不存在显著的正相关关系(r=0.0925,P=0.6931)。我们推断人类活动的干扰和生境的片断化是导致思茅木姜子濒危现状的主要因素。考虑到目前其遗传多样性水平虽然很高,但各居群个体数量很少,因此应该对思茅木姜子各居群的所有个体实施及时的就地保护;而遗传变异大部分存在于居群内的个体间,所以在迁地保护时应在各居群内大量采样。     

山东地区盐碱土花生种子际土壤微生物群落结构的研究

【目的】以不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤为实验对象,探讨花生种子在吸水膨胀与萌发过程中,不同类型盐碱土对种子际土壤微生物多样性变化的影响。【方法】采集不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤,通过对各样品中细菌的16S r RNA基因的V3-V4区进行PCR扩增,利用Illumina Hiseq高通量测序技术对12份V3-V4高变区PCR产物进行测序,并对测序数据进行生物信息学分析。【结果】(1)盐碱土壤的种子际细菌群落多样性高于非盐碱土壤,且以东营青坨滨海盐土种子际土壤细菌群落多样性较高。(2)不同类型土壤样本微生物群落结构在纲水平存在明显差异。4种土壤类型种子际土壤细菌共分属于6个菌纲,分别为Proteobacteria、Actinobacteria、Actinobacteria、Bacteroidetes、Acidobacteria和Firmicutes菌纲,并均以Proteobacteria和Actinobacteria菌纲为主要菌纲。全样本菌落结构分析结果表明,4种类型土壤中不同吸胀时间内种子际微生物菌落在门、属水平上的类型和丰度差异最为显著(P0.05)。(3)beta多样性分析和各样本遗传距离(phylogenetic distances)聚类树图分析表明,4个土壤类型的12个土壤样本种子际土壤中微生物群落均可聚为2大类。【结论】土壤含盐量越高其种子际土壤细菌群落多样性较高。不同类型土壤样本微生物群落结构在纲水平存在明显差异,以Proteobacteria和Actinobacteria菌纲为主要菌纲。种子吸胀萌发时间影响种子际微生物菌落在门、属水平上的类型和丰度,但对相同土壤类型样本间遗传距离无影响。     

基于宏微观结合的甾体微生物羟化反应中菌体形态的表征

为考察生长调节剂对甾体微生物转化过程中菌体生长状况的影响,利用沉降密度宏观定量表征菌体的形态和扫描电镜微观观测菌体形态相结合的方法,描述了生长过程中菌体形态的变化。结果表明:与未加调节剂的空白体系对照,加调节剂体系菌体在各个生长时期均呈现出较小的沉降密度。通过SEM图像可以明显看出加调节剂体系菌体表面光滑,长势丰满、挺拔,生长状况优良。两种方法均表明调节剂的加入明显促进了菌体的生长。     

The LEF1/CYLD axis and cIAPs regulate RIP1 deubiquitination and trigger apoptosis in selenite-treated colorectal cancer cells

Inhibitor-of-apoptosis protein (IAP) inhibitors have been reported to synergistically reduce cell viability in combination with a variety of chemotherapeutic drugs via targeted cellular IAP (cIAP) depletion. Here, we found that cIAP silencing sensitised colorectal cancer (CRC) cells to selenite-induced apoptosis. Upon selenite treatment, the K63-linked ubiquitin chains on receptor-interacting protein 1 (RIP1) were removed, leading to the formation of the death-inducing complex and subsequent caspase-8 activation. Although the ubiquitinases cIAP1 and cIAP2 were significantly downregulated after a 24-h selenite treatment, cylindromatosis (CYLD) deubiquitinase protein levels were marginally upregulated. Chromatin immunoprecipitation assays revealed that lymphoid enhancer factor-1 (LEF1) dissociated from the CYLD promoter upon selenite treatment, thus abolishing suppression of CYLD gene expression. We corroborated these findings in a CRC xenograft animal model using immunohistochemistry. Collectively, our findings demonstrate that selenite caused CYLD upregulation via LEF1 and cIAP downregulation, both of which contribute to the degradation of ubiquitin chains on RIP1 and subsequent caspase-8 activation and apoptosis. Importantly, our results identify a LEF1-binding site in the CYLD promoter as a potential target for combinational therapy as an alternative to cIAPs.     

重组人粒系集落刺激因子(rhG-CSF)对白血病细胞HL-60生长的影响

本文研究了rhG CSF对人白血病细胞系HL 6 0的作用。结果表明 :rhG CSF能够显著抑制HL 6 0细胞生长和C myc基因的表达 ,降低3H TdR的摄入。在含rhG CSF的培养液中经过 2～ 5天的培养 ,部分HL 6 0细胞具备NBT还原能力。这或许说明rhG CSF能导致HL 6 0细胞向成熟方向分化的结果。     

腺相关病毒与人多药耐药基因重组载体的构建及在NIH3T3细胞中的表达

腺相关病毒 (adeno- associated virus,AAV)属细小病毒科 ,是一种最小的动物病毒 .具有其他病毒载体所没有的优点 ,在基因治疗中日益受到瞩目 .以 AAV的一种多克隆载体为基础 ,构建了携带 MDR1基因的重组腺相关病毒载体 (r AAV- MDR1 ) ,经 2 93细胞包装成重组病毒 .将重组质粒、重组病毒分别转染和感染 NIH3T3细胞 ,用 PCR和 MTT法检测了人 MDR1基因的转导及表达 .为 MDR1基因用于临床和腺相关病毒载体在基因治疗中的应用提供了依据