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1.
将鹅源腺病毒Y81G4株全基因组DNA的HindⅢ酶切片段分别插入质粒pUC18,成功构建了全基因组DNA文库。在此基础上,将重组质粒携带的插入片段切出、回收并分别用地高辛标记后作为探针,与经限制酶BamHⅠ、EcoRⅠ、PstⅠ、EcoRⅤ消化的病毒基因组DNA进行Southern Blotting,杂交结果经比较综合后获得了该病毒基因组DNA的HindⅢ、BamHⅠ、EcoRⅠ、PstⅠ、Ec  相似文献   

2.
采用密度梯度离心法及RNase消化法制备并纯化了鲤(GyprinuscarpioLinnaeus)肝脏线粒体DNA(mtDNA),用10种限制性内切酶对mtDNA进行了分析,鲤鱼mtDNA分子量约10.12×10 ̄6,约16.49kb.SalⅠ、PstⅠ、BamHⅠ、XbaⅠ、BglⅠ、PvuⅡ、XhoⅠ、EcoRⅠ、DraⅠ和HindⅢ分别为1、1、3、3、3、4、1、4、4、和6个切点。根据单酶解及双酶解结果,构建了鲤mtDNA10种具酶30个切点的限制性酶切图谱。  相似文献   

3.
胡子鲇mtDNA多态性及限制性酶切图谱   总被引:7,自引:1,他引:6  
用8种限制性内切酶对胡子鲇(ClariasfuscusLacepede)肝脏线粒体DNA(MitochondrialDNA,mtDNA)进行了分析。XhoⅠ、EcoRⅠ、PstⅠ、BamHⅠ、XbaⅠ、HindⅢ在mtDNA分子上分别有2、3、1、1、3和5个切点。胡子鲇种内存在mtDNA酶切片段长度多态性(Restrictionfragmentlengthpolymorphisms,RFLP)。经BglⅠ、BglⅡ酶解,mtDNA都出现两种酶切类型,Ⅰ型各具2个片段,Ⅱ型各具1个片段。mtDNA分子量为10.242×106u,长度约为16.68kb。用双酶解法建立了胡子鲇mtDNA的限制性酶切图谱,并对RFLP现象进行了分析。  相似文献   

4.
对蜀伯毒蛾核型多角体病毒(Parocneria orienta Nuclear polyhedrovirus,简称PaorNPV)形态结构、结构多肽、限制内切酶图谱等特性进行了研究。采用不连续系统垂直板SDS-PAGE分析了PaorNPV的多角体蛋白、病毒粒子结构多肽。应用5种限制性内切酶对PaorNPV基因组DNA进行了酶切分析。结果表明:经热处理的多角体蛋白仅有一条带,分子量为31.5kD,不  相似文献   

5.
应用谷实夜蛾核型多角体病毒(HzSNPV)DNA聚合酶基因HindⅢ/PstⅠ3596bp片段作探针,经Sourthernblot杂交,克隆了中国棉铃虫核型多角体病毒(HaSNPV)完整的DNA聚合酶基因,大小约为3.4kb。限制性内切酶分析表明,HaSNPVDNA聚合酶基因限制性内切酶图谱与HzSNPV相似。用双脱氧链终止法测定该基因部分核苷酸序列(805bp),推导出编码区206a。序列同源性比较显示,HaSNPVDNA聚合酶与HzSNPV之间具有高度的同源性;与LdMNPV、AcMNPV、BmSNPV、CfMNPV和OpMNPV也具有一定的同源性  相似文献   

6.
将抗癌胚抗原(CEA)单链抗体基因插入家蚕杆状病毒转移载体pBacPAK-His,与修饰的家蚕核型多角体病毒Bm-BacPAK DNA共转染家蚕细胞,经同源重组得到含有在多角体蛋白基因启动子控制下的抗CEA ScFv基因的重组病毒Bm-BacScFv。用重组病毒分别感染家蚕细胞和幼虫,在两者中均得到了高效表达,产物分子量为28kD,前者占细胞总蛋白的6%,后者为0.3mg/蚕。目的基因在家蚕细胞和  相似文献   

7.
木毒蛾质型多角体病毒呈不规则多面体,大小差异较大,大多数在0.8-2.1μm之间;病毒粒子在多角体内随机分布,病毒粒子为正二十面体结构,有12个顶体,每个顶上均有一个球状结构,平均直径约为60nm。经SDS-PAGE分析,多角体蛋白由一条分子量为31kD之间。多角体蛋白富含天冬氨酸,谷氨酸及酪氨酸,而半胱氨酸和甲硫氨酸含量较少,其碱性氨基酸与酸性氨基酸之比为1.43。病毒核酸对RNase和DNas  相似文献   

8.
山羊品种间线粒体DNA限制性片段长度多态性研究   总被引:26,自引:2,他引:24  
本试验利用ApaⅠ、BamHⅠ、BclⅠ、BglⅠ、BglⅡ、ClaⅠ、EcoRⅠ、EcoRⅤ、HaeⅡ、HindⅢ、KpnⅠPstⅠ、PvuⅡ、SacⅠ、SalⅠ、SmaⅠ和XhoⅠ计18种限制性内切酶,研究了来自欧洲,非洲及国内的5个山羊品种共计33只个体的mtDNA,共检测了27处限制性态型,可归结为8种单倍型。结果表明,国内2个百品种的基本单倍型为BamHⅠ-A,BclⅠ-B、ClaⅠ-A  相似文献   

9.
隼形目鹰科11种鸟类线粒体DNA分子进化的研究   总被引:6,自引:0,他引:6  
采用ApaⅠ,BamHⅠ,BalⅡ,EcoRⅠ,EcoR V,HindⅢ,HpaⅠ,KpnⅠ,PstⅠ,PvuⅡ,SalⅠ,ScaⅠ,XbaⅠ和XhoⅠ等14种限制性内切酶,对隼形目鹰科11种鸟类(金雕、乌雕、草原雕、普通狂、大狂、鹊、白尾鹞、乌灰鹞、黑翅鸢、高山兀鹫)线粒体DNA限制性片段长度多态性分析。结果表明:是遗传距离最小的是金雕和草原雕(P=0.960),最大的是金雕和兀鹫(P=19.1  相似文献   

10.
将抗癌胚抗原(CEA)单链抗体基因插入家蚕杆状病毒转移载体pBacPAKHis, 与修饰的家蚕核型多角体病毒BmBacPAKDNA共转染家蚕细胞, 经同源重组得到含有在多角体蛋白基因启动子控制下的抗CEAScFv 基因的重组病毒BmBacScFv。用重组病毒分别感染家蚕细胞和幼虫, 在两者中均得到了高效表达, 产物分子量为28kD, 前者占细胞总蛋白的6 % , 后者为0 .3 mg/ 蚕。目的基因在家蚕细胞和幼虫中表达产物经Ni2+IDASepharose6B亲和柱纯化, 前者纯度可达90% 以上, 后者纯度较低; 纯化后的融合蛋白具有CEA 结合活力, 其亲和常数分别为5 .4×108/mol·L- 1 和2.3 ×108/mol·L-1 , 略低于其亲本单抗E7B10 2.7 ×109/mol·L- 1 。  相似文献   

11.
通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.  相似文献   

12.
Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m−2 s−1. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to Pfr.  相似文献   

13.
The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.  相似文献   

14.
A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.  相似文献   

15.
龙胆科药用植物化学成分的研究现状   总被引:16,自引:0,他引:16  
龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。  相似文献   

16.
17.
Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H2-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H2 with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD+, FMN+, and NAD(P)+ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H2OCO2+2H++2e- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation  相似文献   

18.
Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)8 barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)8 structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive ET and active ER. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive ET and active ER represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.  相似文献   

19.
20.
(鱼句)亚科花(鱼骨)型鱼类骨骼系统的比较   总被引:2,自引:0,他引:2  
对我国花型Hemibarbuspattern鱼类作了骨骼系统比较,结果表明,此类型鱼类脑颅较长,副蝶骨平直或稍弯曲,眶蝶骨腹纵嵴发达(铜鱼Coreius septentrionalis例外),下颞窝和咽突中等大,基枕骨后突发达;脑颅中的上筛骨的后突、侧筛骨的外筛突,蝶耳骨的外突、上耳骨的后突、围眶骨和后颞窝等均有明显的差异;咽颅中的舌颌骨、尾舌骨、鳃盖骨和下咽齿的列数等又有显著的区别;附肢骨骼中的腰带骨、脊椎骨中的复合神经骨和第4椎骨腹侧的悬器等也有不同之处。据此,这些差异和区别可作为属间或种间的分类依据。  相似文献   

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