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1.
用双脱氧核苷酸研究了大肠杆菌DNA聚合酶I的Klenow片段的“碱基选择”作用。当引物末端为3'OH时,Mn^2+代替Mg^2+对dNTP的Km影响不大,而使ddNTP的Ki大大降低,非配对的ddNTP仍无抑制作用。当引物末端为ddNMP且与模板配对时是DNA合成的抑制剂,与模板配对的下一个dNTP可增强这一抑制作用,Mn^2+代替Mg^2+使dNTP的诱导作用大大增强而ddNTP无诱导作用。这些  相似文献   

2.
随机扩增多态DNA影响因素的研究   总被引:76,自引:0,他引:76  
随机拉多态DNA分析受诸多因素的影响,我们发现不同厂家制造的PCR扩增仪,不同厂家出品的TaqDNA聚合酶和PCR缓冲液,RAPD反应体系中的引物浓度,Mg^2+浓度,dNTP浓度,BSA和明胶,以及模板DNA的量等均可能对RAPD结果有不同程度的影响。  相似文献   

3.
测定了RB69 DNA 聚合酶以不正确的核苷酸(rNTP、ddNTP以及碱基不配对的dNTP)为底物进行聚合反应的稳态动力学常数,并与Klenow 酶进行了比较.结果表明,RB69 DNA 聚合酶在以不正确的核苷酸为底物进行聚合反应时,其Km 值与正确底物参入时相比有大幅度提高,而kcat保持不变或下降幅度较小.而Klenow 酶在利用不正确的核苷酸为底物时,与正确底物参入时相比,其kcat大幅度下降,而Km (或KD)基本保持不变或上升较小幅度.两种酶不同的动力学特点反映出它们不同的底物选择机制.  相似文献   

4.
扩增条件对茶类植物RAPD带的影响   总被引:15,自引:0,他引:15  
采用梯度分析的方法试验了模板DNA、引物、镁离子、dNTP和Taq酶的浓度对茶类植物进行RAPD分析中DNA扩增结果的影响。实验表明这些条件的变化对扩增出来的RAPD带的数目和强弱会产生影响。经过比较分析,筛选出对于茶类植物进行RAPD分析较理想的扩增条件:2.0mmol/LMgCl2,200umol/LdNTP,15ng引物/20ul反应体积,4ng模板DNA/ul反应体积,1UTaq酶/20ul反应体积。  相似文献   

5.
用电泳迁移分析方法研究了21nt脱氧寡核苷酸G3TG2TGT2G5TG2TGT(CP1)与129bp的乙肝病毒(HBV)核衣壳启动子(Cp)片段内一位点结合形成的三链DNA的特异性及稳定性.在克隆有HBV基因组的质粒pCP10的酶切产物中,CP1仅与含Cp的129bp片段结合.在20mmol/LMg2+溶液中其解离常数(Kd)为1.4×10-7mol/L.不同离子稳定三链DNA的效果依次为sp4+(精胺)>Mg2+>Zn2+>Na+>K+,离子之间存在相互竞争作用.比CP1多一误配碱基的脱氧寡核苷酸G2TG2TGTG3TG2TG2TG2T(CP2)在20mmol/LMg2+溶液中与Cp结合的Kd值约为CP1的1/7,而在60mmol/LK+或5mmol/LZn2+溶液中检测不到它与Cp的结合,这进一步显示了三链DNA形成的特异性.细胞的生理离子浓度被认为是:Sp4+1mmol/L,Mg2+10mmol/L,K+140mmol/L,因此,CP1在细胞内将能特异地与Cp结合并具有较好的稳定性.  相似文献   

6.
用Ca2+和胰酶处理大叶藻(Zosteramarina)和刺松藻(Codiumfragile)叶绿体膜,研究了它们的类囊体膜多肽组分与Mg2+诱导Chla荧光和膜表面电荷变化之间的相互关系。观察到:(1)在大叶藻的叶绿体膜中,Mg2+诱导PS-Ⅱ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;但这种相关性的效应不存在于刺松藻的叶绿体膜中。(2)用Ca2+处理这两种叶绿体膜分别除去其类囊体膜表面的32-34KD和30-31KD多肽,对上述Mg2+诱导的现象无明显的影响。(3)用胰酶进一步消化这些Ca2+处理过的叶绿体膜分别除去其类囊体膜表面的26KD和23-24KD多肽,那么在大叶藻的叶绿体膜中,Mg2+诱导荧光和类囊体膜表面电荷变化的相关性效应则全部消失;但在刺松藻的叶绿体膜中,Mg2+诱导荧光增高的效应完全消失,而Mg2+诱导膜表面电荷变化的性质则保持不变。这些实验结果不仅证明,类囊体膜表面的26KD和23-24KD多肽分别为大叶藻和刺松藻叶绿体膜中阳离子诱导激发能在PS-Ⅱ和PS-Ⅰ之间分配变化的特异性作用部位;而且说明阳离子调节激发能在这两个光系统之间分配的机理,在这两种海生植物的叶绿体膜中  相似文献   

7.
几种因素对山茶属植物RAPD分析的DNA扩增的影响   总被引:10,自引:0,他引:10  
唐绍清  施苏华  林海波   《广西植物》1998,18(2):185-188
多种因素会影响RAPD扩增,本研究试验了引物、Mg2+和dNTP的浓度以及Taq酶来源对山茶属植物进行RAPD分析的DNA扩增的影响。结果表明这些因素对扩增结果都会产生影响,通过比较分析,得到了一个对于山茶属植物进行RAPD分析较理想的扩增条件。  相似文献   

8.
宁晓檬  赵晓岩 《病毒学报》1996,12(4):355-359
提取感染鸡胚成纤维细胞MDV-I弱毒株814病毒DNA为模板,根据RBIB株gB基因5′及3′两端核苷酸离列设计引物,利用PCR技术扩增了我国MDV-I弱毒株814gB基因(2.9kb)将扩增片段平末端克隆到载体pBluescriptSK中EcoRV位点,经BamHI,HindIII酶切鉴定得到不同插入方向的重组质粒。构建圹增片段的酶切图谱及部分序列分析证明与RBIB株gB基因无差异,显示了极高的  相似文献   

9.
彩绒革盖菌多酚氧化酶活性研究   总被引:12,自引:1,他引:11  
本文测定了彩绒革盖菌在PDY液体培养基中的多酚氧化酶活性。在30℃,110r/min,恒温振荡培养条件下,多酚氧化酶第14天达产酶高峰,最高酶活549.0u;酶作用的最适酸碱度为pH5.0;最适作用温度30℃;Mn2+、Ba2+、Mg2+等离子对多酚氧化酶有激活作用,Ag+、Fe3+等离子对酶活则有明显的抑制作用  相似文献   

10.
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   

11.
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13.
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   

14.
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   

15.
16.
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   

17.
18.
For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.  相似文献   

19.
Å. Berg 《Bird Study》2013,60(2):153-165
CapsuleThe amount of forest (at local and landscape scales) and occurrence of residual habitats at the local scale are shown to be the major factors influencing bird community composition in farmland–forest landscapes in central Sweden.

Aims To investigate the importance of local habitat and landscape structure for breeding birds in farmland–forest landscapes in central Sweden.

Methods Breeding birds were censused at 292 points. A detailed habitat mapping was made within 300 m of the points. Within a 300–600 m radius only two major habitats (forests and arable fields) were identified.

Results Cluster analyses of bird communities identified three site types that also differed in habitat composition: (i) partially forested sites in forested landscapes; (ii) heterogeneous sites with residual habitats in mosaic landscapes; and (iii) field-dominated farmland sites in open landscapes. A total of 19 of 25 farmland bird species (restricted to farmland or using both farmland and forest) had the highest abundance in farmland sites with mosaics of forest and farmland, while only six farmland species had the highest abundance in field-dominated sites. The bird community changed from being dominated by farmland species to being dominated by forest species (common in forest landscapes without farmland) at small proportions (10–20%) of forest at the local scale. A major difference in habitat composition between heterogeneous and field-dominated sites was the occurrence of different residual habitats (e.g. shrubby areas and seminatural grasslands). These habitats seemed to influence bird community composition more than land-use, despite covering <10% of the area. Seminatural grasslands were important for bird community composition and species-richness, but grazing seemed to be less important. Among different land-use types, cereal crops were the least preferred fields. Set-asides with tall vegetation and short rotation coppices were positively associated with species-richness of farmland birds.

Conclusion In general, the composition of the landscape was important for bird community composition, although amount and distribution of forests, occurrence of residual habitats and land-use of fields at the local scale had the strongest influence on bird community composition. The possible implications of these patterns for managing farmland–forest landscapes are discussed.  相似文献   

20.
This work describes the isolation and characterization of twenty-nine compounds from the fruits of Rhus typhina L., including eleven flavonoids (1–11), eleven phenols (12–22), two pentacyclic triterpenes (23–24), two organic acids (25–26), one lumichrome (27), one courmarin (28) and one pyrimidine (29) on the basis of their spectroscopic data. Compounds apigenin (1), daidzein (4), orobol (5), 3′, 5, 5′, 7-tetrahydroxyflavanone (6), naringenin (7), butein (8), (-)-catechin (9), quercetin-3-O-α-L-(3″-O-galloyl)-rhamnoside (11), 2-hydroxybenzoic acid (13), 4-hydroxybenzaldehyde (14), vanillin (15), methyl 3,4-dihydroxybenzoate (16), 3,5-dihydroxybenzamide (18), tyrosol (19), caffeic acid (20), 3-(2,4,6-trihydroxyphenyl)-1-(4-hydroxyphenyl)-propan-1-one (21), phlorizin (22), friedelin (23), oleanolic acid (24), 4,4-dimethyl-heptanedioic acid (25), anthranilic acid (26), lumichrome (27), scoparone (28) and uracil (29) have not been recorded before in this plant. This is the first report on the occurrence of compounds 4–7, 9, 11, 13–14, 16, 18–21, 25–29 from the genus Rhus. Moreover, the chemotaxonomic significance of these isolated compounds was also summarized.  相似文献   

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