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91.
地衣芽孢杆菌β-甘露聚糖酶的纯化及酶学性质 总被引:6,自引:0,他引:6
地衣芽孢杆菌(Bacilluslicheniformis)NK-27菌株发酵产生的β-甘露聚糖酶(β-mannanase)经硫酸铵盐析沉淀,两次DEAE纤维素和SephadexG-100离子交换柱层析以及制备PAGE筹步骤,获得了凝胶电泳均一的样品。用SDS-凝胶电泳测得纯化后的β-甘露聚糖酶分子量为26kD,用凝胶聚焦电泳测得等电点PI为5.0。酶反应的最适pH为9.0,最后温度为60℃,稳定pH为6.0—9.0,稳定温度为40℃。金属离子中Mg ̄(2+)、Ca ̄(2+)、Fe ̄(2+)、Ni ̄(2+)对该酶有一定的激活作用;而Sn ̄(2+)、Zn ̄(2+)、Al ̄(3+)、Ag ̄+和Hg ̄(2+)对该酶有强烈的抑制作用。NK-27菌株的β-甘露聚糖酶对魔芋葡萄甘露聚糖和角豆胶半乳甘露聚糖的Km值分别为7.14和5.56mg·ml ̄(-1);V_(max)分别为200.53和157.45μmol·mg ̄(-1)·min ̄(-1)。 相似文献
92.
Potent inhibition of human immunodeficiency virus type 1 in primary T cells and alveolar macrophages by a combination anti-Rev strategy delivered in an adeno-associated virus vector. 总被引:1,自引:0,他引:1 下载免费PDF全文
R T Inouye B Du D Boldt-Houle A Ferrante I W Park S M Hammer L Duan J E Groopman R J Pomerantz E F Terwilliger 《Journal of virology》1997,71(5):4071-4078
The rate of viral replication appears to play a pivotal role in human immunodeficiency virus type 1 (HIV-1) pathogenesis and disease progression as it outstrips the capacity of the immune system to respond. Important cellular sites for HIV-1 production include T lymphocytes and tissue macrophages. Antiviral strategies, including newer treatment modalities such as gene therapy of HIV-1-susceptible cell populations, must be capable of engendering durable inhibitory effects to HIV-1 replication in both of these primary cell types in order to be effective. Among the potential genetic targets for intervention in the HIV-1 life cycle, the Rev regulatory system, consisting of Rev and its binding site, the Rev-responsive element (RRE), stands out as particularly attractive. Rev is essential for maintaining the stability of the viral genomic RNA as well as viral mRNAs encoding key structural and regulatory proteins. Moreover, it exhibits favorable threshold kinetics, in that Rev concentrations must rise above a critical level to exert their effect. To disable Rev function, primary T cells or macrophages were transduced with anti-Rev single-chain immunoglobulin (SFv) or RRE decoy genes either singly or in combination by employing adeno-associated virus vectors and then challenged with HIV-1. By directing both a protein and a nucleic acid against the normal interaction between Rev and the RRE, this genetic antiviral strategy effectively inhibited infection by either clinical or laboratory virus isolates. These results provide a framework for novel interventions to reduce virus production in the infected host. 相似文献
93.
Adeno-associated virus type 2-mediated transfer of ecotropic retrovirus receptor cDNA allows ecotropic retroviral transduction of established and primary human cells. 总被引:1,自引:0,他引:1 下载免费PDF全文
K Qing T Bachelot P Mukherjee X S Wang L Peng M C Yoder P Leboulch A Srivastava 《Journal of virology》1997,71(7):5663-5667
The cellular receptors that mediate binding and internalization of retroviruses have recently been identified. The concentration and accessibility of these receptors are critical determinants in accomplishing successful gene transfer with retrovirus-based vectors. Murine retroviruses containing ecotropic glycoproteins do not infect human cells since human cells do not express the receptor that binds the ecotropic glycoproteins. To enable human cells to become permissive for ecotropic retrovirus-mediated gene transfer, we have developed a recombinant adeno-associated virus type 2 (AAV) vector containing ecotropic retroviral receptor (ecoR) cDNA under the control of the Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter (vRSVp-ecoR). Established human cell lines, such as HeLa and KB, known to be nonpermissive for murine ecotropic retroviruses, became permissive for infection by a retroviral vector containing a bacterial gene for resistance to neomycin (RV-Neo(r)), with a transduction efficiency of up to 47%, following transduction with vRSVp-ecoR, as determined by the development of colonies that were resistant to the drug G418, a neomycin analog. No G418-resistant colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. Southern and Northern blot analyses revealed stable integration and long-term expression, respectively, of the transduced murine ecoR gene in clonal isolates of HeLa and KB cells. Similarly, ecotropic retrovirus-mediated Neo(r) transduction of primary human CD34+ hematopoietic progenitor cells from normal bone marrow was also documented, but only following infection with vRSVp-ecoR. The retroviral transduction efficiency was approximately 7% without prestimulation and approximately 14% with prestimulation of CD34+ cells with cytokines, as determined by hematopoietic clonogenic assays. No G418-resistant progenitor cell colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. These results suggest that sequential transduction of primary human cells with two different viral vectors may overcome limitations encountered with a single vector. Thus, the combined use of AAV- and retrovirus-based vectors may have important clinical implications for ex vivo and in vivo human gene therapy. 相似文献
94.
Augmented sympathetic tone alters muscle metabolism with exercise: lack of evidence for functional sympatholysis 总被引:2,自引:0,他引:2
Shoemaker J. Kevin; Pandey Prasant; Herr Michael D.; Silber David H.; Yang Qing X.; Smith Michael B.; Gray Kristen; Sinoway Lawrence I. 《Journal of applied physiology》1997,82(6):1932-1938
Shoemaker, J. Kevin, Prasant Pandey, Michael D. Herr, DavidH. Silber, Qing X. Yang, Michael B. Smith, Kristen Gray, and LawrenceI. Sinoway. Augmented sympathetic tone alters muscle metabolismwith exercise: lack of evidence for functional sympatholysis. J. Appl. Physiol. 82(6):1932-1938, 1997.It is unclear whether sympathetic tone opposesdilator influences in exercising skeletal muscle. We examined highlevels of sympathetic tone, evoked by lower body negative pressure(LBNP, 60 mmHg) on intramuscular pH and phosphocreatine (PCr)levels (31P-nuclear magnetic resonance spectroscopy) duringgraded rhythmic handgrip (30 contractions/min; ~17, 34, 52 and 69%maximal voluntary contraction). Exercise was performedwith LBNP and without LBNP (Control). At the end of exercise, LBNPcaused lower levels of muscle pH (6.59 ± 0.09) comparedwith Control (6.78 ± 0.05; P < 0.05). PCr recovery, an index of mitochondrial respiration, was lessduring the recovery phase of the LBNP trial. Exercise mean arterialpressure was not altered by LBNP. The protocols were repeated withmeasurements of forearm blood flow velocity and deep venous samples(active forearm) of hemoglobin (Hb) saturation, pH, and lactate. WithLBNP, mean blood velocity was reduced at rest, during exercise, andduring recovery compared with Control (P < 0.05). Also, venous Hbsaturation and pH levels during exercise and recovery were lower withLBNP and lactate was higher compared with Control(P < 0.05). We concludethat LBNP enhanced sympathetic tone and reduced oxygen transport. Athigh workloads, there was a greater reliance on nonoxidativemetabolism. In other words, sympatholysis did not occur. 相似文献
95.
阳离子诱导大叶藻叶绿体膜中激发能在PSⅡ和PSⅠ之间分配变化的机理(英文) 总被引:1,自引:0,他引:1
用Ca2+ 和胰酶处理大叶藻(Zostera m arina)叶绿体膜研究了其类囊体膜多肽成分与Mg2+ 诱导其Chla荧光和类囊体膜表面电荷变化之间的相互关系,观察到:1.在正常的叶绿体膜中,Mg2+ 诱导PSⅡ荧光强度的增高与其诱导类囊体膜表面电荷密度的降低密切相关;2.用Ca2+ 处理这种叶绿体膜,除去类囊体膜表面的32~34 kD多肽对Mg2+ 诱导的上述现象无影响;3.如果用胰酶消化Ca2+ 处理过的叶绿体膜,进一步除去膜表面的26 kD多肽,Mg2+诱导的这些现象则全部消失。这些实验结果清楚地表明,在大叶藻的叶绿体膜中,类囊体膜表面的26 kD 多肽是阳离子诱导这两种相关现象的特异性作用部位。对阳离子调节激发能在PSⅡ和PSⅠ之间分配的机理进行了讨论 相似文献
96.
Qing Li Gerhard W. Hacker Gorm Danscher Ursula Sonnleitner-Wittauer Lars Grimelius 《Histochemistry and cell biology》1995,104(2):145-150
Silver staining techniques developed to demonstrate argyrophilic nucleolar organizer regions (Ag-NORs) have been widely applied in a variety of cell kinetic studies, using the mean number of AgNORs in tumour cells as a marker for malignancy of certain types of neoplasms. However, the AgNOR techniques currently available are not entirely satisfactory, as unspecific silver precipitates readily form in the sections. On the other hand, the contrast staining, may be so weak as to render identification of the AgNORs difficult. In the present study, some of the key factors influencing the outcome of AgNOR staining were evaluated in a more systematic way. A modified AgNOR staining procedure is now proposed, giving highly contrasting AgNORs with minimal unspecific silver precipitation, thus facilitating both manual and computerized counting. The new technique involves the use of microwave irradiation in order to shorten the processing time, the use of gelatin as a protective colloid, and a Farmer's solution to optimize the specificity of the technique. 相似文献
97.
98.
无性系植物种群生态学研究进展及有关概念 总被引:74,自引:5,他引:69
无性系植物种群生态学研究进展及有关概念刘庆钟章成(中国科学院成都生物研究所,610041)(西南师范大学生物系,重庆630715)AdvancesinEcologicalResearchofClonalPlantPopulationandSomeRe... 相似文献
99.
1987年9月至1989年8月,笔者对来自云南瑞丽的2只雄性马来熊成体和7只幼体(3♂,4♀♀)在人工饲养条件下进行了观察研究,本文报道在不同生长时期的不同饲料配方、幼熊感觉器官的发育、齿式的形成、外部形态的变化、体重的生长方法及行为。 相似文献
100.
急性病毒性肝炎的发病率慢性肝炎患病率和肝病死亡率的研究 总被引:15,自引:0,他引:15
1984~1987年,在黑龙江、河北、河南、湖南、上海五个省市城乡10.08855人口中进行急性肝炎发病率、慢性肝炎患病率、与病毒性肝炎有关的肝病死亡率的研究。急性肝炎标化发病率为152.19/10万,主要发生在20~50岁组人群;因无甲肝暴发流行,除上海外各点季节发病率分布均衡。慢性肝炎标化患病率为158.25/10万(诊断标准为6个月前有明确急性肝炎病史,现有明显的临床症状或体征,肝功能异常,故实际慢肝患病率要高于此数字);与病毒性肝炎有关的肝病死亡(包括肝癌)标化率为22.65/10万,其中肝病为 13.14/10万。男性死亡率显著高于女性。 相似文献