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The studies of affectors affecting RAPD were carried out by the 12 species of CLanatus as template DNAThe results showed that the purification of template DNA,the concentrations of template DNA,Taq polymerase,random primer,Mg2+and the PCR programe played an important role in RAPD analysisBased on the researches above,a technical system for RAPD analysis in species of Clanatus was established:in 10 L reaction solution,containing 10 ng/10 L template DNA,05 mol/L random primer,50 mmol/L TrisHCl(pH83),500... The studies of affectors affecting RAPD were carried out by the 12 species of CLanatus as template DNAThe results showed that the purification of template DNA,the concentrations of template DNA,Taq polymerase,random primer,Mg2+and the PCR programe played an important role in RAPD analysisBased on the researches above,a technical system for RAPD analysis in species of Clanatus was established:in 10 L reaction solution,containing 10 ng/10 L template DNA,05 mol/L random primer,50 mmol/L TrisHCl(pH83),500 g/mL BSA,25 mmol/L MgCl2,06 U/10 L Taq DNA,200 mol/L dNTPs 以西瓜种内的12个品种(系)为模板DNA,研究了影响RAPD扩增效果的因素。结果表明,模板DNA、Taq酶、随机引物、Mg2+浓度以及PCR反应程序在RAPD分析中都具有重要的作用。根据以上因子的研究结果,建立了适合西瓜种植物的RAPD分析的技术体系:10μL反应液中,模板10ng/10μL,引物05μmol/L,TrisHCl(pH83)50mmol/L,BSA500μg/mL,MgCl225mmol/L,TaqDNA06U/10μL,dNTPs200μmol/L,10%蔗糖400和1mmol/L甲酚红。 Contraceptive levonorgestrel (LNG) was coupled to polyα、β(3hydroxypropyl)DLaspartamide,a biodegradable and biocompatible material.The drug loaded in the polymer was about 27.0% (w/w). The polymer conjugate was characterized by differential scanning calorimertry (DSC) and fourier transform infrared (FTIR). The in vitro release of levonorgestrel from the biocompatible polymer conjugate was studied in TrisHCl (0.1 mol/L) and trypsin. Levonorgertrel was measured by HPLC. The data indicate that this levonorgestrel... Contraceptive levonorgestrel (LNG) was coupled to polyα、β(3hydroxypropyl)DLaspartamide,a biodegradable and biocompatible material.The drug loaded in the polymer was about 27.0% (w/w). The polymer conjugate was characterized by differential scanning calorimertry (DSC) and fourier transform infrared (FTIR). The in vitro release of levonorgestrel from the biocompatible polymer conjugate was studied in TrisHCl (0.1 mol/L) and trypsin. Levonorgertrel was measured by HPLC. The data indicate that this levonorgestrel polymer conjugate can be released in these two systems but faster in trypsin.$ 具有临床意义的甾体类避孕药左旋 1 8甲基炔诺酮 (L NG)键合于生物相容性聚氨基酸材料 :α、β 聚 (3羟丙基 ) DL 天冬酰胺上 ,制成长效控释药物。用红外光谱法、差热分析等分别对材料及键合药物进行了表征 ,药物的接入率达 2 7.0 % (w/ w) (n=3) ,共价缩合物以棒状形式释药进行了体外水解和酶解释放试验。结果表明 ,酶解释放比水解释放略快 A proteinase,which has important work on turbot digestive,was purified from turbot(Scophthalmus maximus L.) and its major physical and chemical characteristics were studied and reported in this article.The intestine proteinase extraction was prepared through homogenate and centrifugation by Tris-HCl buffer at low temperature.The supernatant was purified through ammonium sulfate grading precipitation further and the active component was dialyzed and concentrated.Then the condensed proteinase was purified by means... A proteinase,which has important work on turbot digestive,was purified from turbot(Scophthalmus maximus L.) and its major physical and chemical characteristics were studied and reported in this article.The intestine proteinase extraction was prepared through homogenate and centrifugation by Tris-HCl buffer at low temperature.The supernatant was purified through ammonium sulfate grading precipitation further and the active component was dialyzed and concentrated.Then the condensed proteinase was purified by means of DEAE-Sepharose Fast Flow anion-exchange chromatography with an elution of a linear gradient of 5 mmol·L~(-1) Tris-HCl buffer(pH 8.5) containing 1.0 mol·L~(-1) NaCl.At last,the pure enzyme was obtained through Sephadex G-100 chromatography with an elution of 5mmol·L~(1) TrisHCl buffer(pH 8.5).The purity of the purified intestine proteinase was confirmed by the presence of a single band on SDS-PAGE.The relative molecular mass of this enzyme was determined to be about 58 kD.By using casein as substrate to measure proteinase activity,the characterization of turbot intestine proteinase was made.The enzyme is stable at pH 6.0-11.0 and temperature below 30 ℃.When the temperature rose,there was a rapid decline of its stability.The enzyme showed maximum activity at pH 9.0 and 50 ℃.When the temperature declined,the optimum pH of enzyme showed a trend of moving to alkaline.And when the action time prolonged,the optimum temperature of enzyme showed a trend to decline.Furthermore,the enzyme is activated by Mn~(2+) and inactivated by Cu~(2+),Zn~(2+),Ag~+,Fe~(3+).In further studies,it was inhibited by SH-enzyme inhibitors such as p-Hydroxymercuribenzoate(PCMB) and N-bromosuccinimide(NBS) remarkably,and partially inhibited by tosyl-lysine chloromethyl ketone(TLCK).Using casein as the substrate, the Km value of enzyme is 2.92 g·L~(-1) at pH 9.0,50 ℃.The result showed the enzyme seemed to be a SH-enzyme. 以大菱鲆肠道为材料,对大菱鲆消化生理中起重要作用的肠蛋白酶的分离纯化条件和理化性质进行了研究。经Tris-HCl缓冲液抽提,硫酸铵分级分离,DEAE-Sepharose Fast Flow离子交换层析、Sephadex G-100凝胶过滤层析分离纯化,获得大菱鲆肠蛋白酶Ⅰ的电泳纯制品,并对该酶的性质进行了研究。结果显示:纯酶的分子量约为58kD。纯酶最适反应pH为9.0,最适反应温度50℃;pH稳定范围6.0~11.0,30℃以下酶活性稳定;Mn2+可激活大菱鲆肠蛋白酶Ⅰ,Cu2+、Zn2+、Ag+、Fe3+则对酶活性有抑制作用;巯基蛋白酶抑制剂显著抑制大菱鲆肠蛋白酶Ⅰ的活性,金属蛋白酶抑制剂对大菱鲆肠蛋白酶Ⅰ无影响。50℃、pH9.0条件下以双倒数作图法得大菱鲆肠蛋白酶Ⅰ催化酪蛋白水解的Km值为2.92g.L-1。
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