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1.
菜豆幼苗EPSP合成酶的分离纯化和它的部分性质   总被引:3,自引:0,他引:3  
利用硫酸铵分级沉淀,Sephedex G-50凝胶柱层析,FPLC Mono-Q和磷酸纤维素离子层析法从菜豆幼苗中分离提纯了EPSP合成酶。该酶被纯化2961.6倍,比活性达到6219.4nmolmg^-1蛋白min^-1。该酶分子量经SDS-PAGE检测为51kD,等电点为pH5.7,酶促反应最适pH7.5,最适温度45℃。6.2μmol/L的除草剂草甘膦能抑制EPSP合成酶活性的50%。  相似文献   

2.
巨大芽孢杆菌产胞外青霉素酰化酶发酵液经硫酸铵分级抽提及SephadexG-100、羟基磷灰石、DEAE纤维素DE52等层析步骤,提纯了青霉素酰化酶,得到电泳均一的酶制剂。纯酶比活力约为25U/mg蛋白,纯化49倍,活力回收58%,经PAGE及SDS-PAGE测知该酶不含亚基,其分子量约为140kD。该酶最适pH为9.0,最适温度47℃,用底物NIPAB测活,其Km值为6.2×10~(-4)mol/L,Vm值为1.24×104mol/L。此外还探讨了部分金属离子对该酶的影响。  相似文献   

3.
亲和层析纯化HepG2细胞分泌的PAI—1   总被引:1,自引:0,他引:1  
本文报道用抗PAI-1单克隆抗体(McAb)亲和层析建立了纯化PAI-1的简便方法。经免疫亲和层析,Sephacryl S200凝胶过滤,从HepG2细胞培液中分离到糖基化和非糖基化两种形式的PAI-1,回收率为84%,PAI-1比活性为6.1×10^4IU/mg。糖基化PAI-1分子量为50kD,比活性5.8×10^4IU/mg。非糖基化PAI-1分子量43kD,占总PAI-130%,仍具有PA  相似文献   

4.
本文报道用抗PAI-1单克隆抗体(McAb)亲和层析建立了纯化PAI-1的简便方法。经免疫亲和层析,SephacrylS200凝胶过滤,从HepG2细胞培液中分离到糖基化和非糖基化两种形式的PAI-1,回收率为84%,PAI-1比活性6.1×104IU/mg。糖基化PAI-1分子量为50kD,比活性5.8×104IU/mg。非糖基化PAI-1分子量43kD,占总PAI-130%,仍具有PAI-1活性。用ConA-Sepharose亲和层析进一步纯化得到SDS-PAGE纯的糖基化PAI-1。  相似文献   

5.
利用DPA使Tb3+的荧先强度显著增强的原理进行带3蛋白活性的测定,方法简便、灵敏、重复性好,而且能够进行连续荧光扫描测量.应用连续荧光扫描法测定了带3蛋白介导的DPA与Cl-交换的动力学特征参数.结果表明,带3蛋白介导的DPA与Cl-的交换对DIDS非常敏感,受DIDS的强烈抑制,抑制程度大于90%;DPA由内向外转运的米氏常Km=28.1—31.2mmol/L;带3蛋白的天然底物Cl-从内侧竞争性抑制DPA向外转运,抑制常数ki=60.4±6.9mmol/L;膜内侧DPA与外侧Cl-交换的活化能,在4—25℃范围内为5.8±0.5kCal/mol25—37℃范围内为19.8±1.5kCal/mol;膜内侧DPA与外侧Cl-交换受转运介质pH(膜内外对称改变)的显著影响,pH<7.4时,交换速度显著升高.本实验证明DPA确是经带3蛋白而转运的,但转运机制可能与无机离子转运有所不同。  相似文献   

6.
P物质对GABAA和GABAB受体介导的DRG神经元膜反应的调制作用   总被引:10,自引:2,他引:8  
关兵才  李之望 《生理学报》1994,46(5):441-450
实验在幼年大鼠DRG标本上进行。应用细胞内记录观察了SP对GABA反应的调制作用。结果证明:(1)单独滴加SP(5×10(-6)-4×10(-5)mol/L)或浴槽灌流SP(10(-6)-5×10(-6)mol/L)不引起膜电位的改变或仅有轻微的去极化,但却能使GABA引起的去极化反应减小50.8±20.2%(±SD)(20/30);(2)单独滴加SP可使多数受检细胞APD50延长28.7±9.1%(±SD)(10/18);(3)在预加SP后,能使baclofen所引起的APD50缩短效应(20.6±2.9%,±SD)完全消除(4/12)或翻转成APD(50)延长19.3±8.9%(±SD)(8/12);(4)预加GABAB受体激动剂baclofen(10(-4)-10(-3)mol/L)30—90s后明显地抑制muscimol(10-4-10-3mol/L)引起的去极化反应,其抑制效应达54.4±18.8%(±SD)(17/20)。由于DRG神经元的胞体通常可用来作为研究初级传入终末的模型,因而本文实验结果提示:介导伤害性刺激信息的P物质在背角的释放,可能作用于初级传入终末,从而产生对抗GABA介导的突触  相似文献   

7.
铝对玉米生长和硝酸还原酶活性的影响   总被引:2,自引:0,他引:2  
测定了生长在Al2(SO4)3100μmol/L氮素营养液中的两个玉米品种(SC704和VA35的根系和叶片)的NADH-硝酸还原酶(EC1.6.6.1)和NAD(P)H-硝酸还原酶(EC1.6.6.2)活性。  相似文献   

8.
首次报道了人肝脏血红素加氧酶的同工酶的分离纯化,并初步探讨了它们的性质,采用DEAE-SephadexA-25和羟基磷灰石柱层析法从人肝脏分离纯化HO的同工酶,酶活性检测、SDS-PAGE分析结果显示,人肝脏微粒体含量HO的同工酶,按洗脱先后顺序分别得到分子量为30000和36000的HO-1和HO-2.酶促反应中需相同辅酶参与,其中酶活性HO-1明显高于HO-2,两之比为2.4:1,从分子量和  相似文献   

9.
高效价甘薯羽状斑驳病毒抗血清的制备   总被引:6,自引:0,他引:6  
用嫁接方法将甘薯羽状斑驳病毒(SPFMV)接种到I.setosa上扩繁,以0.2mol/LpH7.2PBK缓冲液、垫层差速离心、蔗糖密度梯度离心提取纯化SPFMV。纯化的SPFMVOD260/280的比值为1.25。将纯化的SPFMV免疫家兔制备抗血清,在环状沉淀和微量沉淀试验中,用提纯病毒测定抗血清的效价均为1:4096;以SPFMV-IgG为第一抗体,应用Dot-ELISA对甘薯和I.selosa叶片中的SPFMV分别作了测定。  相似文献   

10.
蚯蚓纤溶酶的纯化及稳定性研究   总被引:4,自引:0,他引:4  
蚯蚓纤溶酶(EPA)抽提液经30% ~70% 饱和度的(NH4)2SO4 盐析、DEAE—纤维素柱层析、Sephadex G-75葡聚糖凝胶过滤等纯化步骤,得到了具有纤溶活性的洗脱峰,置凝胶电泳后,得到四个活性组分,它们经50℃保温6h,活力上升64% ;在2 m ol/L盐酸胍存在时,活力仅保存7.2% ,当其浓度降低时,活力可恢复至90% ;在1% SDS存在时,活力仅保存12.1% ,但当SDS除去时,活力又可恢复。因此,盐酸胍、SDS均为EPA 可逆性抑制剂。另外,EPA 中含有较高的糖链(占总量的45% ),具有良好的抵抗自水解作用。  相似文献   

11.
Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.  相似文献   

12.
正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases  相似文献   

13.
The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.  相似文献   

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17.
鸡传染性法氏囊病病毒研究进展   总被引:3,自引:0,他引:3  
闫笑  李天宪 《中国病毒学》2003,18(2):191-195
传染性法氏囊病(Infection bursal disease, IBD)是由鸡传染性法氏囊病毒(Infectious bursal disease virus, IBDV)引起的鸡和火鸡的一种高度接触性传染病,给世界各国的禽养殖业带来了巨大损失.自IBDV发现至今新的变异株不断出现,分子结构的改变导致病毒致病力的改变及宿主对疫苗应答的改变,使得传统的疫苗已不能控制其流行,因此各国学者对其基因组结构和功能进行了广泛深入的研究,并积极研制新型有效的疫苗以达到防治的目的.  相似文献   

18.
In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.  相似文献   

19.
Shen  Jia-Yuan  Li  Man  Xie  Lyu  Mao  Jia-Rong  Zhou  Hong-Ning  Wang  Pei-Gang  Jiang  Jin-Yong  An  Jing 《中国病毒学》2021,36(1):145-148
正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).  相似文献   

20.
Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.  相似文献   

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