温度对海带幼孢子体生长和光合作用的影响

根据欧盟项目的总体研究计划 ,在室内和围隔实验中研究了海带幼孢子体在不同温度条件下的生长情况和光合作用曲线 ,得出幼孢子体在 15°C下生长情况最好 ,而且其长度和重量的日增加量均和温度有线性相关关系。 Pm 值在 15℃时为 5 79mol O2 / (g DW· h) ,围隔实验中为 6 4 2 (mol O2 / (g DW· h)。由于围隔实验海区的光照强于室内 ,使得围隔中海带的光合作用曲线其光饱和点高于室内实验海带的光饱和点。海带光饱和参数 (Ik)与温度有线性相关关系 (R2 =0 .736 7,p<0 .0 5 )。5℃时 ,Ik 值平均为 96μE/ (m2 · s) ,10℃时为 71μE/ (m2 · s) ,15℃时为 31μE/ (m2 · s)。呼吸速率 R值在 5℃时最低 ,为 5 4μmol O2 / (g DW·h) ,10℃、15℃和围隔中均高于 5℃时的呼吸速率 ,并且在 10℃达到最高。本次实验研究得出的生长和光合作用参数将有助于合理确定桑沟湾中海带的养殖容量。     

《Bergey''''s Manual of Systematic Bacteriology》

Bergey’s Manual of Systematic Bacteriology(伯吉氏系统细菌学手册)第一卷最近由美国Williams & Wilkins公司出版。R.G.E.Murray为本书编辑委员会主任委员。本书由不同国度     

用PCR检测细胞培养中支原体污染

细胞培养中支原体污染已经成为严重的问题.为了扩增6种支原体(精氨酸支原体,口腔支原体,人型支原体,猪鼻支原体,发酵支原体及莱氏支原体)核糖体RNA操纵子的16s和23s DNA间区,设计了三个通用PCR引物(F1,F2及R1).当以6种支原体DNA为模板时,引物F1和R1产生340到468bp的片段,引物F2和R1产生145到211bp的片段,当用Hela细胞或E.coli DNA作为模板,用引物F1和R1时,在电泳中未观察到特定区带.此法最小能检出8.5fg精氨酸支原体DNA,相当于13个精氨酸支原体.这说明,当这些支原体污染细胞培养时,能用PCR法检测出来.     

群体分布对夏大豆产量和水分利用效率的影响

在田间试验条件下研究了群体不同分布对夏大豆产量构成和水分利用的影响.夏大豆‘鲁豆4号'(Glycine max cv. Ludou 4)在同一群体密度(3.09×10~5株/hm~2)下设5种分布方式,即行距×株距分别为A:18 cm×18 cm,B:27 cm×12 cm,C:36 cm×9 cm,D:45 cm×7.2 cm,E:54 cm×6cm.结果表明,群体分布影响夏大豆的产量、叶片水分特征和水分利用效率(WUE).A、B处理的产量显著高于D、E处理(P<0.05),其他处理间均无显著差异;随着行距加大,单株有效荚数、粒数及百粒重呈下降趋势;叶片相对含水量(RWC)、水势(Ψw)和渗透势(Ψs)随生育进程的推进呈整体下降趋势,其中,A、B处理RWC、Ψw 和Ψs的平均值均显著高于D、E处理,E处理的Ψw在日变化的正午阶段明显低于其他处理;WUE与行距呈负相关(R=-0.935~*),与产量呈正相关(R=0.997~(**)),其中,A、B处理的WUE显著高于其他处理(P<0.05),D、E处理极显著低于B处理(P<0.01).夏大豆植株相对均匀分布的处理可改善产量构成因素及叶片水分状况,进而形成较高的产量,提高水分利用效率.     

ERIC-PCR在人工混合菌体系中的检出灵敏度*

提取大肠杆菌DH5α和阴沟肠杆菌 (E 2 6R)的基因组DNA ,进行ERIC PCR ,可以分别得到稳定而独特的DNA指纹图谱 ;模板量的变动范围从 1 0～ 1 0 0ng都不会影响其图谱的稳定性 ;在图谱中DH5α和E 2 6R各自均有一条含量最大的特征带 ,其不易受实验环境和实验条件的干扰而发生变化。把DH5α和E 2 6R按比例混合 ,提取混合菌基因组DNA ,进行ERIC PCR ,结果表明 :混合菌的DNA指纹图谱为各纯菌图谱的叠加 ,亦具有稳定性 ;通过对凝胶电泳图谱中特征带的分析 ,可     

一种新的以细胞表面受体为靶向的基因导入系统

鉴于胰岛素样生长因子Ⅰ号及Ⅱ号的受体 (IGFⅠR ,IGFⅡR)在人原发性肝癌中过量表达 ,以及表皮细胞生长因子受体 (EGFR)在多种人恶性肿瘤过量表达 ,设计和合成了针对IGFⅠR及IGFⅡR的 1 4肽E5和针对EGFR的 1 6肽GE7,以及流感病毒血凝素功能域 2 0肽HA2 0作为内吞小体释放寡肽 (Endosomereleasingoligopeptide,EOP) ,将它们分别与多聚阳离子多肽 (Polycationic polypeptide ,PCP)———多聚赖氨酸 (Polylysine ,PL)或鱼精蛋白 (Protamine,PA)共价连接 ,藉静电效应与DNA形成一个复合体 (E5 PCP/DNA/PCP HA2 0 ,GE7 PCP/DNA/PCP HA2 0 ) ,即构建的新的受体介导的靶向性非病毒型基因导入系统 .体内、外实验结果表明它们相对靶向且高效地将外源基因导入人恶性肿瘤细胞并得到预期表达     

大肠杆菌质粒突变体(p#GTES)的特性分析

大肠杆菌(E.coli)RRI质粒(pGTE5)的β-内酰胺酶基因(β-lactamas gene)可以在E. coli与枯草杆菌(Bacillus subtilis)中复制和表达。但是,在E. coli细胞中向胞外分泌β-内酰胺酶量少,活力低,对氨苄青霉素(Ampicillin,简称Ap)的抗性也低。将该菌株用紫外线(30w,40cm,150s)和硫酸二乙酯双重诱变,在含Ap的LB平板上筛选到已．Coli质粒突变体(p#GTE5)。     

E75、R78和D82是影响大肠埃希氏菌FtsZ自身组装及其与MreB相互作用的重要氨基酸

【目的】探索大肠埃希氏菌Escherichia coli FtsZ突变体FtsZ~(E75A)、FtsZ~(R78G)和FtsZ~(D82A)对FtsZ自身组装和FtsZ-MreB相互作用的影响。【方法】利用常规分子克隆和定点突变技术,构建FtsZ及其突变体表达载体,亲和纯化得到相应的目标蛋白;通过同源重组构建QN6(ftsZ::yfp-cat)、QN7(ftsZ~(E75A)::yfp-cat)、QN8(ftsZ~(R78G)::yfp-cat)和QN9(ftsZ~(D82A)::yfp-cat)菌株;利用活细胞成像技术观察FtsZ及其突变体的胞内定位模式;免疫沉淀和细菌双杂交实验检测FtsZ/FtsZ*-FtsZ*或FtsZ/FtsZ*-MreB间的相互作用;光扫描检测定点突变对FtsZ组装特性的影响。【结果】FtsZ~(E75A)、FtsZ~(R78G)和FtsZ~(D82A)突变体的功能活性降低、各突变体在E.coli内不能正确的定位和形成功能性Z环;FtsZ/FtsZ*-FtsZ*单体间的相互作用减弱或消失,FtsZ*-MreB相互作用破坏;FtsZ突变体体外聚合效率降低。【结论】FtsZ E75、R78和D82是影响FtsZ正确组装和功能及FtsZ-MreB相互作用的重要氨基酸。     

在大气干旱条件下胀果甘草气孔振荡的RLC电路模拟

建立了植物水分输导和蒸腾的电阻(R)、电感(L)、电容(C)电路模型。实验测定出的胀果甘草气孔振荡发生、持续和衰减的阈值与模型分析结果一致。当蒸腾拉力(F)大于输导阻力(R)时气孔开始振荡并使振幅逐渐加大;当F=R时气孔振荡的振幅和频率不变;当0<(R-F)<2(1/2)/(L/C)时气孔振荡开始减弱;当(R-F)>2(1/2)/(L/C)时气孔不会出现振荡。在本实验条件下胀果甘草的R=3.1×10~9MPa·m~(-3)·s,2(1/2)/(L/C)=3.11-3.64×10~5MPa·m~(-3)·s。     

产过敏素harpin的固氮工程菌的某些特性研究*

研究了产过敏素harpin的固氮工程菌(Enterobacter cloacae E4)在番茄、烟草叶片上的致过敏能力及该菌所携的双质粒的稳定性。试验结果表明:E4与DH5(pCPP430)致过敏能力的速度和强度基本相同。E4与308R(pCPP430)相比,烟草上它们致过敏能力的速度基本一致,但308R(pCPP430)致过敏能力的强度更强,在番茄叶片上,E4和308R(pCPP430)致过敏能力的速度和强度基本一样。E4所携的双质粒pCPP430和pMC73A在宿主细菌中是不稳定的,在宿     

Poly(4-thiouridylic acid) as messenger RNA and its application for photoaffinity labelling of the ribosomal mRNA binding site.

Poly(4-thiouridylic acid) [poly(s4U)] synthesized by polymerization of 4-thiouridine 5'-diphosphate with Escherichia coli polynucleotide phosphorylase (EC 2.7.7.8) acts as messenger RNA in vitro in a protein-synthesizing system from E. coli. It stimulates binding of Phe-tRNA to ribosomes both in the presence of EF-Tu-Ts at 5 mM Mg2+ concentration and nonenzymatically at 20 mM Mg2+ concentration. It codes for the synthesis of polyphenylalanine. Poly(s4U) competes with poly(U) for binding to E. coli ribosomes. Light of 330 nm photoactivates poly(s4U) thus making it a useful photoaffinity label for the ribosomal mRNA binding site. Upon irradiation of 70-S ribosomal complexes, photoreaction occurs with ribosomal proteins as well as 16-S RNA. Ribosomes pre-incubated with R17 RNA are protected against the photoaffinity reaction. The labelling of 16-S RNA can be reduced by treatment of ribosomes with colicin E3.     

Ca2+ sensitization and potentiation of the maximum level of myofibrillar ATPase activity caused by mutations of troponin T found in familial hypertrophic cardiomyopathy

Human wild-type cardiac troponin T, I, C and five troponin T mutants (I79N, R92Q, F110I, E244D, and R278C) causing familial hypertrophic cardiomyopathy were expressed in Escherichia coli, and then were purified and incorporated into rabbit cardiac myofibrils using a troponin exchange technique. The Ca2+-sensitive ATPase activity of these myofibrillar preparations was measured in order to examine the functional consequences of these troponin mutations. An I79N troponin T mutation was found to cause a definite increase in Ca2+ sensitivity of the myofibrillar ATPase activity without inducing any significant change in the maximum level of ATPase activity. A detailed analysis indicated the inhibitory action of troponin I to be impaired by the I79N troponin T mutation. Two more troponin T mutations (R92Q and R278C) were also found to have a Ca2+-sensitizing effect without inducing any change in maximum ATPase activity. Two other troponin T mutations (F110I and E244D) had no Ca2+-sensitizing effects on the ATPase activity, but remarkably potentiated the maximum level of ATPase activity. These findings indicate that hypertrophic cardiomyopathy-linked troponin T mutations have at least two different effects on the Ca2+-sensitive ATPase activity, Ca2+-sensitization and potentiation of the maximum level of the ATPase activity.     

PEGylation of a proprotein convertase peptide inhibitor for vaginal route of drug delivery: In vitro bioactivity,stability and in vivo pharmacokinetics

Uterine proprotein convertase (PC) 6 is critical for embryo implantation in mice and women. It is also one of the PC family members that play a vital role in HIV infectivity. We hypothesized that inhibiting PC6 in the female reproductive tract (vagina, cervix and uterus), may protect women from both pregnancy and HIV infection. One key requirement to prove this concept in an animal model is a vaginally deliverable PC6 inhibitor. Nona-d-arginine (Poly R) is a potent peptide PC inhibitor and is able to inhibit HIV in cell culture. We modified Poly R by PEGylation with different strategies and determined their biochemical properties in vitro and in vivo. PEGylation at the C-terminus, regardless of the PEG size (30 kDa or 1239 Da) did not compromise the inhibitory potency of Poly R. In contrast, PEGylation at both termini (1239 Da) dramatically reduced its inhibitory activity. Poly R and C-PEGylated Poly Rs also showed equal potency in inhibiting a PC6-dependent cellular process critical for embryo implantation. Poly R and the equipotent C-PEGylated Poly Rs were further tested for their serum stability in vitro and pharmacokinetics in vivo following vaginal administration in mice. All Poly Rs were equally stable in mouse serum in vitro for 24 h; C-PEGylated Poly Rs showed enhanced vaginal absorption and penetration across the vaginal mucosa/epithelium. This is the first report that C-terminal PEGylation significantly enhances the therapeutic properties of Poly R for vaginal drug delivery. Our findings also provide important insights into future design of Poly R derivatives.     

Induction of interferon by polyribonucleotides containing thiopyrimidines.

The influence of thioketo substitution in pyrimidine bases of double-stranded polynucleotides on interferon induction was investigated. The stabilizing effect of 2-thioketo substitution was reflected in the increased interferon inducing activity of poly(A-s2U) over that of poly(A-U). Poly(A-s2U) and poly(I)-poly(s2C) were as effective as poly(I)-Poly(C) in rabbit cells. Poly(I)-poly(C) and poly(I)-poly(s2C) were compared in several animal species. No differences in biological effects were observed in rabbits and dogs. In rodents, poly(I)-poly(s2C) was less effective and less toxic.Poly(I)-poly(s2C) was highly resistant against degradation by human serum. Further investigations seem to be justified to elucidate whether this property offers any advantages for the potential clinical utilization of poly(I)-poly(s2C).     

牛心朴子须根的化学成分研究

从采自宁夏的萝摩科鹅绒藤属植物牛心朴子 (CynanchumkomaroviiAl.Iljinski.)须根的乙醇提取物中分离鉴定了十个非C2 1 甾体类化合物 :β D 呋喃果糖基 (2→ 1) α D [6 O 芥子酰基 ] 吡喃葡萄糖甙 (1) ,β D (3 O 芥子酰基 ) 呋喃果糖基 (2→ 1) α D [6 O 芥子酰基 ] 吡喃葡萄糖甙 (2 ) ,[6 O β D 吡喃葡萄糖基 (1→ 6 ) β D 吡喃葡萄糖基 1,2 双氧 (4 羟基 3,5 二甲氧基肉桂酰 ) (3) ,7 脱甲氧基娃儿藤碱 (4) ,9 羟基 芳樟醇 3 O β D 吡喃木糖基 (1→ 6 ) β D 吡喃葡萄糖甙 (5 ) ,(2E ,6R) 2 ,6 二甲基 2 ,7 辛二烯 1,6 二醇 (6 ) ,[(+) 丁香素 ](7) ,4′ O demethylepiyangambin(8) ,4′ 羟基 2′ 甲氧基苯乙酮 (9) ,(2S ,3S ,4R ,12E) N [2′ (R) 羟基二十二碳烷基 ] 1,3,4 三羟基 2 酰胺 二十碳烷基 12 烯 (10 )。除化合物 4和 9外 ,其余化合物均为首次从该植物中分离得到。     

双酚A诱导雄性中国林蛙肝细胞雌激素受体表达和卵黄蛋白原合成

为探讨双酚A(bisphenol A,BPA)对雄性两栖动物肝细胞中雌激素受体(estrogenreceptor,ER)表达和卵黄蛋白原(vitellogenin,VTG)合成的影响,将中国林蛙(Ranachensinensis)雄性成体分别持续暴露于10-7、10-6、10-5mol/LBPA水体中10、20、30d,设10-9、10-8mol/L雌二醇(E2)为阳性对照。用原位杂交技术检测ERmRNA在肝细胞中的表达定位,用免疫组织化学技术检测肝细胞中ER和VTG蛋白的表达。结果显示,各BPA和E2处理组肝细胞中均有ERmRNA阳性反应,ER和VTG蛋白的表达相对值均显著高于空白对照组。在同一暴露时间,ER和VTG表达值是随着BPA浓度的增加呈增高趋势。在同一BPA浓度,VTG表达值是随暴露时间的延长呈增高趋势,而ER表达值则无显著性变化。这些表明BPA可以通过诱导雄性中国林蛙肝细胞的ER高调导致VTG合成,但其效应远低于E2。     

大肠杆菌N~α-乙酰基转移酶RimI对人胸腺素α原N末端乙酰化修饰的影响

目的:研究大肠杆菌Nα-乙酰基转移酶RimI对人胸腺素α原(ProTα)N末端乙酰化修饰的影响。方法:在大肠杆菌中共表达ProTα与大肠杆菌Nα-乙酰基转移酶RimI,同时设置只表达ProTα的对照菌,分别提取纯化ProTα,利用HPLC检测其发生乙酰化修饰的程度。结果:ProTα的乙酰化修饰发生在翻译后水平,有无RimI的过表达、诱导时间的长短以及这两个因素之间的交叉效应均对ProTα的乙酰化程度有影响,其F值分别为53.8、89.22及16.28,P值均小于0.0001;无论有无RimI过表达,乙酰化的ProTα所占比例在诱导6 h后逐渐升高(P<0.0001);在过表达Ri-mI的菌株中,乙酰化的ProTα在诱导12 h后占37.4%,而在无RimI过表达的菌株中占64.3%;RimI对ProTα乙酰化的抑制作用从诱导6 h后开始显现(P=0.0007)。结论:在大肠杆菌中,过量的Nα-乙酰基转移酶RimI可抑制人胸腺素α原的乙酰化修饰。     

Latch and trigger role for R445 in DAT transport explains molecular basis of DTDS

A recent study reports on five different mutations as sources of dopamine transporter (DAT) deficiency syndrome (DTDS). One of these mutations, R445C, is believed to be located on the intracellular side of DAT distal to the primary (S1) or secondary (S2) sites to which substrate binding is understood to occur. Thus, the molecular mechanism by which the R445C mutation results in DAT transport deficiency has eluded explanation. However, the recently reported X-ray structures of the endogenous amine transporters for dDAT and hSERT revealed the presence of a putative salt bridge between R445 and E428 suggesting a possible mechanism. To evaluate whether the R445C effect is a result of a salt bridge interaction, the mutants R445E, E428R, and the double mutant E428R/R445E were generated. The single mutants R445E and E428R displayed loss of binding and transport properties of the substrate [³H]DA and inhibitor [³H]CFT at the cell surface while the double mutant E428R/R445E, although nonfunctional, restored [³H]DA and [³H]CFT binding affinity to that of WT. Structure based analyses of these results led to a model wherein R445 plays a dual role in normal DAT function. R445 acts as a component of a latch in its formation of a salt bridge with E428 which holds the primary substrate binding site (S1) in place and helps enforce the inward closed protein state. When this salt bridge is broken, R445 acts as a trigger which disrupts a local polar network and leads to the release of the N-terminus from its position inducing the inward closed state to one allowing the inward open state. In this manner, both the loss of binding and transport properties of the R445C variant are explained.     

蜜蜂化学生态学化学通讯与信息素研究进展

概括了蜜蜂化学生态学的主要研究内容和领域,重点评述了蜂群内的化学通讯和蜜蜂信息素研究的进展。迄今,已经鉴定的蜂王信息素有９－氧化－（反）－２－癸烯酸（９－ＯＤＡ）,Ｒ（一）－９－羟基癸烯酸Ｒ（一）－９ＨＤＡ）,Ｓ（＋）－９－羟基癸烯酸（Ｓ（＋）－９ＨＤＡ）,对－羟基苯甲酸甲酯（ＨＯＢ）和４－羟基－３－甲本基乙醇（ＨＶＡ）等５种;工蜂那氏信息互膛牛醇,橙花醇,（反,反）－法尼醇,（反）柠檬醛,（顺）