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101.
籼稻232蜡质基因转录起始位点的鉴定   总被引:5,自引:0,他引:5  
高继平  郦永忠 《遗传学报》1995,22(6):431-436
Northernblot杂交分析和蜡质基因cDNA的序列分析表明水稻蜡质基因的转录本可能延伸到翻译起始密码子(ATG)上游12kb处。据此设计了21Nt的寡核苷酸引物,并以籼稻232胚乳RNA为模板,以引物延伸法确定籼稻232蜡质基因的转录起始点,籼稻蜡质基因的转录起始旁邻顺序CTCACCA与高等植物基因的转录起始点一致顺序CTCATCA仅相差1个碱基。通过顺序比较,对东乡野生稻蜡质基因中的转录起始位点的位置,以及对此两稻种中TATA盒的可能顺序进行了讨论。  相似文献   
102.
乙肝病毒PreSl片段与乙肝表面抗原羧端的融合表达   总被引:4,自引:0,他引:4  
我们利用聚合酶链反应法(PCR)得到了编码乙肝病毒肝细胞受体结合位点PreSl(21 ̄47)的基因片段,并将它分别融合到S基因中相应于第175,188和223位氨基酸残基处。所得到的融合基因插入痘苗病毒表达载体pGJP-5后,在哺乳动物细胞CV-1中进行了暂时表达,对融合蛋白的表达、分泌和抗原性的研究表明,3种融合基因均能表达具有S和PreSl双重抗原性的融合蛋白,但融合位点对表达水平和分泌性质有  相似文献   
103.
病毒基因组启动子识别的人工神经网络方法   总被引:1,自引:0,他引:1  
本文运用神经网络方法,并结合病毒基因分子生物学有关理论与统计事实,对病毒基因启动子区域进行了识别,文中选择了共35个基因组,作为研究对象.学习组选择了28个基因组,预测组选择了7个基因组,结果表明,将神经网络模型与病毒基因有关理论相结合,能够运用计算方法,以大量的可能启动子组合中排列出唯一的启动子区域.  相似文献   
104.
In a previous report [Yanget al., (1987a),J. Biol Chem. 262, 7034–7040], a cyclic-AMP- and calcium-independent brain kinase which requires autophosphorylation for activity was identified as a very potent myelin basic protein (MBP) kinase. In this report, the phosphorylation sites of MBP by this autophosphorylation-dependent protein kinase (autokinase) are further determined by two-dimensional electrophoresis/thin-layer chromatography, phosphoamino acid analysis, high-performance liquid chromatography, tryptic peptide mapping, sequential manual Edman degradation, and direct peptide sequencing. Autokinase phosphorylates MBP on both threonine and serine residues. Three major tryptic phosphopeptide peaks were resolved by C18-reversed phase highper-formance liquid chromatography. Sequential manual Edman degradation together with direct sequence analysis revealed that FS(p)WGAEGQKPGFGYGGR is the phosphorylation site sequence (molar ratio 1.0) for the first major phosphopeptide peak. When mapping with bovine brain MBP sequence, we finally demonstrate Ser115, one of thein vivo phosphorylation sites in MBP, as the major site phosphorylated by autokinase, implicating a physiologically relevant role of autokinase in the regulation of brain myelin function. By using the same approach, we also identified HRDT(p)GILDSLGR (molar ratio 0.9) and TT(p)HYGSLPQK (molar ratio 0.8) as the major phosphorylation site sequences in32P-MBP phosphorylated by autokinase, further indicating that -Arg-XSer/Thr-(neutral amino acid)3-(amino acid-containing hydroxyl group such as Ser/Glu/Asp)-(neutral amino acid)2-may represent a unique consensus sequence motif specifically recognized by this autophosphorylation-dependent multisubstrate/ multifunctional protein kinase in the brain.  相似文献   
105.
A quantitative endonuclease assay, which relies on the introduction of single and double strand breaks into supercoiled plasmid DNA, was used to study the activity of the extracellular nuclease of Serratia marcescens SM6 in buffer and in groundwater. The parallel enzyme concentration-dependent production of relaxed and linear plasmid molecules suggests that the nuclease produces single and double strand breaks in duplex DNA. Bovine serum albumin stimulated the nuclease activity towards DNA and RNA and increased the stability of the enzyme against thermal inactivation. The DNase activity at 4 °C and 50 °C was almost half of that at the optimum temperature (37 °C). The nuclease was active in groundwater, although the specific activity was lower than in buffer. In a groundwater aquifer microcosm, mineral-adsorbed transforming DNA was substantially less accessible to the nuclease than was dissolved DNA. The data suggest that the extracellular nuclease of Serratia marcescens may contribute to DNA turnover in the environment and that adsorption of DNA to minerals provides protection against the nuclease.Abbreviations GW groundwater GWA groundwater aquifer  相似文献   
106.
UV irradiation of the ATPase (CF1) from spinach chloroplasts in the presence of 3'-arylazido-β-alanyl-8-azido ATP (8,3'-DiN3ATP) results in a nucleotide-dependent inactivation of the enzyme and in a nucleotide-dependent formation of -β cross-links. The results demonstrate an interfacial localization of the nucleotide binding sites on CF1.  相似文献   
107.
Claudia Kluge  Peter Dimroth   《FEBS letters》1994,340(3):245-248
Subunit c of the F1F0-ATPase from Propionigenium modestum was extracted from the particulate cell fraction with chloroform/methanol. The protein was further purified by carboxymethyl cellulose chromatography and anion exchange HPLC in the organic solvent. SDS-PAGE of the purified protein indicated a single stained protein band migrating as expected for the c-subunit. Incubation of isolated subunit c in chlorform/methanol or aqueous buffer containing dodecyl-β- -maltoside with [14C]dicyclohexylcarbodiimide (DCCD) resulted in the incorporation of radioactivity into the protein. The rate of this reaction depended on the external pH; it was significantly faster in the more acidic than in the alkaline pH range. In the presence of Na+ subunit c was partially protected from labeling with [14C]DCCD at pH 6.1 and at pH 7.5, whereas no protection was evident at pH 5.5. At pH 7.5, the rate of subunit c labeling by [14C]DCCD in the presence of 20 mM NaCl was about 50% lower than in the absence of Na+ ions. The isolated c-subunit therefore apparently retains in part the Na+ binding site which, when occupied, diminishes the reactivity of the protein towards DCCD.  相似文献   
108.
X-Ray analysis of the ferritin of Escherichia coli (Ec-FTN) and of Ec-FTN crystals soaked in (NH4)2Fe(SO4)2 has revealed the presence of three iron-binding sites per subunit. Two of these form a di-iron site in the centre of the subunit as has been proposed for the ‘ferroxidase centres’ of human ferritin H chains. This di-iron site, lying within the 4-alpha-helix bundle, resemble those of ribonucleotide reductase, methane monoxygenase and haemerythrin. The third iron is bound by ligands unique to Ec-FTN on the inner surface of the protein shell. It is speculated that this state may represent the nucleation centre of a novel type of Fe(III) cluster, recently observed in Ec-FTN.  相似文献   
109.
The nest site characteristics of the freshwater three-spined stickleback, Gasterosteus aculeatus (form leiurus ), were quantitatively investigated in springs and the main stream of the Yamayoke and the Tsuya River system, central Japan. Most nests (93·4%) were on a muddy or sandy substratum, at depths of 10–40 cm (84·3%), in water velocities less than 15 cm s−1 (76·2%) and in the temperature range of 14 to 16° C (82·7%), Spring-fed localities provided more of these conditions than the main stream channel and hence contained more potential nesting areas. Thus, they were utilized by male sticklebacks both temporally (prolonged breeding season) and spatially (more nest sites).  相似文献   
110.
Leucrose formation from sucrose and fructose by dextransucrase is of practical interest. It has been investigated at different experimental conditions, including the influence of temperature on reaction rate and selectivity. Under appropriate conditions high product yield can be obtained. Furthermore, a model is presented that allows interpretation of the experimental data.  相似文献   
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