A组轮状病毒SA11VP6基因的克隆和表达

从ＳＡ１１ＶＰ６基因全序列克隆开始,设计一对两端带有酶切位点的引物,逆转录ＰＣＲ扩增出ＶＰ６全基因ＣＤＮＡ。经酶切后插入ＰＵＣ１９,构建了ＶＰ６全基因克隆ＰＲＡ６。再经酶切后插入痘苗病毒载休质凿ＰＪＳＡ１１７５中。利用Ｌｉｐｏｆｅｃｔｉｎ导入ＴＫ１４３细胞,利用ＴＫ基因和Ｌａｃ基因作为重组病毒的筛选标记。表达产物用单克隆抗体ＥＬＩＳＡ法检测,发现细胞培养上清和细胞裂解液都是阳性。Ｗｅｓｔｅｒｎ ｂ     

新疆呼图壁盐化草甸群落的DCA, CCA及DCCA分析

 本文应用DCA、CCA及DCCA排序技术对新疆呼图壁盐化草甸群落进行了分析。分析中选取了地下水位、粘土层出现深度、粘土层厚度、地下水pH值及地下水矿化度5个环境因子;同时为了分析空间格局对植被分异影响的大小,建立了样地空间坐标矩阵。结果表明：地下水位和地下水的pH值是引起植被分异的两个主要因素,空间格局对植被分异的影响大于环境因子对植被分异的影响。     

温度对黄粉虫成虫繁殖的影响

黄粉虫是多种小型经济动物的常用优良饵料,本试验在20.1℃,24.0℃、28.5℃、31.7℃和36.5五种恒温下饲养该成虫结果,成虫寿命平均分别为63.0、54.2、38.8、38.0和26.1天;每雌平均产卵量则分别为200.3、207.3、122.3、115.8和81.2粒,成虫平均生产1g卵消耗麸皮量分别为2.66、2.00、2.19、2.14和4.15g。结果说明人工繁殖黄粉虫的成虫期温度以24℃为最适宜。     

诺卡氏菌属GS-17耐热茁霉多糖酶的提纯和性质

诺卡氏菌属GS-17(Nocardia sp.GS-17)的耐热茁霉多糖酶(Pullulanase EC.3.2.1.41)的粗酶液经中空纤维柱超滤浓缩、羟基磷灰石柱层析和Pullulan-Sepharose 6B亲和层析,得到凝胶电泳均一的纯酶,比活提高264倍.酶作用最适温度为55℃,最适PH6.2,分子量140000,等电点pI为6.0.该酶水解茁霉多糖、支链淀粉和可溶性淀粉,但不水解糖原.酶在50℃作用于茁霉多糖的米氏常数K_m为0.90mg/ml,最大反应速度V_(max)为57μmol·min~(-1)·mg~(-1).Zn~(2 )、Fe~(3 )、Hg~(2 )、Cu~(2 )、Pb~(2 )和环状糊精对酶有抑制作用,Ca~(2 )对酶有激活作用.经蛋白质侧链化学修饰研究表明,色氨酸残基位于酶的活性位区.该酶是由1129个氨基酸残基组成的单肽链,酶的N末端序列经测定为:Ala-Gly-His-Gly-Pro-Asp-Val-Gln-Asp-Gly-     

Uniparental Mitochondrial Transmission in the Cultivated Button Mushroom, Agaricus bisporus

A uniparental mitochondrial (mt) transmission pattern has been previously observed in laboratory matings of the cultivated mushroom Agaricus bisporus on petri dishes. In this study, four sets of specific matings were further examined by taking mycelial plugs from the confluent zone of mated homokaryons and inoculating these plugs into rye grain for laboratory fruiting and for fruiting under industrial conditions. Examination of the mt genotype of each individual fruit body for mt-specific restriction fragment length polymorphisms further confirmed that the mt genome was inherited uniparentally. The vegetative radial growth and the fruiting activity of two pairs of intraspecific heterokaryons, each pair carrying the same combination of nuclear genomes but different mt genotypes, were compared. Our results suggested that the mt genotype did not appreciably affect radial growth or fruiting activity. The failure to recover both heterokaryons, each carrying either parental mt genotype in any given cross, therefore clearly indicated that in matings of A. bisporus, the mt genome from one of the parental homokaryons is either selectively excluded in the newly formed heterokaryon or selectively eliminated in the immediate heterokaryotic mitotic progeny of the newly formed heterokaryon.     

Metabolic activation of unsaturated derivatives of valproic acid. Identification of novel glutathione adducts formed through coenzyme A-dependent and -independent processes

The ability of 2-n-propyl-4-pentenoic acid (Δ⁴-VPA) and 2-n-propyl-2(E)-pentenoic acid ([E]-Δ²-VPA), two unsaturated metabolites of valproic acid (VPA), to form reactive intermediates, deplete hepatic glutathione (GSH) and cause accumulation of liver triglycerides was investigated in the rat. With the aid of ionspray liquid chromatography-tandem mass spectrometry (LC-MS/MS), three GSH adducts were detected in the bile of Δ⁴-VPA-treated animals and were identified as 4-hydroxy-5-glutathion-S-yl-VPA-γ-lactone, 5-glutathion-S-yl-(E)-Δ³-VPA and 3-oxo-5-glutathion-S-yl-VPA. A fourth conjugate was identified tentatively as 4-glutathion-S-yl-5-hydroxy-VPA. Quantitative analysis of the corresponding N-acetylcysteine (NAC) conjugates in urine indicated that metabolism of Δ⁴-VPA via the GSH-dependent pathways accounted for approximately 20% of an acute dose (100 mg kg⁻¹ i.p.). In contrast, when rats were given an equivalent dose of (E)-Δ²-VPA, only one GSH adduct (5-glutathion-S-yl-(E)-Δ³-VPA) was detected at low concentrations in bile. In vitro experiments with rat liver mitochondria demonstrated that Δ⁴-VPA undergoes coenzyme A- and ATP-dependent metabolic activation in this organelle via the β-oxidation pathway to intermediates which bind covalently to proteins. When liver homogenates and hepatic mitochondria from rats injected with Δ⁴-VPA, (E)-Δ²-VPA or VPA were analyzed for GSH content, it was found that only Δ⁴-VPA depleted GSH pools significantly. Treatment of rats with Δ⁴-VPA and (to a lesser extent) VPA led to an accumulation of liver triglycerides, whereas (E)-Δ²-VPA had no measurable effect. It is concluded that Δ⁴-VPA undergoes metabolic activation by both microsomal cytochrome P-450-dependent and mitochondrial coenzyme A-dependent processes, and that the resulting electrophilic intermediates, which are trapped in part by GSH, may mediate the hepatotoxic effects of this compound. In contrast, (E)-Δ²-VPA is not transformed to any appreciable extent to reactive metabolites, which thus accounts for the apparent lack of hepatotoxicity of this positional isomer in the rat.     

New killing system controlled by two genes located immediately upstream of the mukB gene in Escherichia coli

The nucleotide sequence was determined of the region upstream of the mukB gene of Escherichia coli. Two new genes were found, designated kicA and kicB (killing of cell); the gene order is kicB-kicA-mukB. Promoter activities were detected in the regions immediately upstream of kicB and kicA, but not in front of mukB. Gene disruption experiments revealed that the kicA disruptant was nonviable, but the kicB-disrupted mutant and the mutant lacking both the kicB and kicA genes were able to grow. When kicA disruptant cells bearing a temperature-sensitive replication plasmid carrying the kicA ⁺ gene were grown at 30° C and then transferred to 42° C, the mutant cells gradually lost colony-forming ability, even in the presence of a mukB ⁺ plasmid. Rates of protein synthesis, but not of RNA or DNA synthesis, fell dramatically during incubation at 42° C. These results suggested that the kicB gene encodes a killing factor and the kicA gene codes for a protein that suppresses the killing function of the kicB gene product. It was also demonstrated that KicA and KicB can function as a post-segregational killing system, when the genes are transferred from the E. coli chromosome onto a plasmid.     

Repeated polyketide synthase modules involved in the biosynthesis of a heptaene macrolide by Streptomyces sp. FR-008

Genes for biosynthesis of a Streptomyces sp. FR-008 heptaene macrolide antibiotic with antifungal and mosquito larvicidal activity were cloned in Escherichia coli using heterologous DNA probes. The cloned genes were implicated in heptaene biosynthiesis by gene replacement. The FR-008 antibiotic contains a 38-membered, poiyketide-derived macrolide ring. Southern hybridization using probes encoding domains of the type i modular erythromycin polyketide synthase (PKS) showed that the Streptomyces sp. FR-008 PKS gene cluster contains repeated sequences spanning c. 105 kb of contiguous DNA; assuming c. 5 kb for each PKS module, this is in striking agreement with the expectation for the 21-step condensation process required for synthesis of the FR-008 carbon chain. The methods developed for transformation and gene replacement in Streptomyces sp. FR-008 make it possible to genetically manipulate polyene macrolide production, and may later lead to the biosynthesis of novel polyene macrolides.     

向日葵幼苗环旋运动的三维轨迹

采用光学显微镜标尺和垂线原理制作的三维空间点测定仪,对向日葵（Ｈｅｌｉａｎｔｈｕｓ ａｎｎｕｕｓＬ．）幼苗的环旋运动进行了连续测量。结果表明：向日葵环旋运动的轨迹有椭圆型、摆动型和不规则型;同一植株在不同生长阶段所表现的环旋运动轨迹不一定相同,同一株龄的不同个体也不一定具有相同的运动轨迹;运动的方向有左旋和右旋,圆周运动光源可以显著地改变运动方向;从三维角度看,在整个下胚轴生长阶段,环旋运动的振幅存在一个由小变大再由大变小的变化规律     

扁圆封印木（相似种）茎干的解剖特征

贵州省水城矿区晚二叠世煤核中扁圆封印木（相似种Ｓｉｇｉｌｌａｒｉａ ｃｆ．ｂｒａｒｄｉｉＢｒｏｎｇｎ．）茎干的主要解剖特征如下：管状中柱,具多边形薄壁细胞组成的髓。初生木质部成环带状,外缘呈规则的齿槽状,向心式发育。次生木质部显束状特征,横切面管胞为方圆至长方形,纵切面为梯状壁增厚,并具流苏纹。射线１—２列细胞宽,数个至十余个细胞高。叶迹起源于初生木质部外缘的槽中,中始式,但以向心发育为主。