酶蛋白

The profile of liver CYP450 induction by DEX in male rat After 4 daily i.p. administering of 0,25, 50 and lOOmg/kg/d DEX in male Wistar rat, it was determined the total CYP450 content, the mRNA and protein expression levels of CYP3A1, CYP3A2 and CYP2B1/2, and the ERD (CYP3A catalytic activity), PROD (CYP2B catalytic activity) and BROD (total CYP450 catalytic activity) in livers.

DEX对雄性大鼠肝脏CYP450的诱导效应雄性Wistar大鼠腹注0、25、50和100mg DEX/kg/d诱导处理4天后，测定大鼠肝脏总CYP450含量，CYP3A1、CYP3A2和CYP2B1/2的mRNA及蛋白表达水平，肝脏红酶素脱甲基酶(ERD，CYP3A活性)，苄氧基试卤灵脱乙基酶(PROD，CYP2B活性)，苯氧基试卤灵脱乙基酶(BROD，总CYP450活性)，结果表明，CYP450含量、ERD、PROD和BROD在DEX多次诱导后都有升高，CYP3A1 mRNA表达水平、蛋白含量和酶活性有明显的升高，剂量效应关系明显；CYP3A2蛋白也有明显升高，而其mRNA表达水平却没有变化，提示CYP3A1和CYP3A2的诱导机制可能不同。

The profile of liver CYP450 induction by DEX in male ratAfter 4 daily i.p. administering of 0,25, 50 and lOOmg/kg/d DEX in male Wistar rat, it was determined the total CYP450 content, the mRNA and protein expression levels of CYP3A1, CYP3A2 and CYP2B1/2, and the ERD (CYP3A catalytic activity), PROD (CYP2B catalytic activity) and BROD (total CYP450 catalytic activity) in livers.

DEX对雄性大鼠肝脏CYP450的诱导效应雄性Wistar大鼠腹注0、25、50和100mg DEX/kg/d诱导处理4天后，测定大鼠肝脏总CYP450含量，CYP3A1、CYP3A2和CYP2B1/2的mRNA及蛋白表达水平，肝脏红酶素脱甲基酶(ERD，CYP3A活性)，苄氧基试卤灵脱乙基酶(PROD，CYP2B活性)，苯氧基试卤灵脱乙基酶(BROD，总CYP450活性)，结果表明，CYP450含量、ERD、PROD和BROD在DEX多次诱导后都有升高，CYP3A1 mRNA表达水平、蛋白含量和酶活性有明显的升高，剂量效应关系明显；CYP3A2蛋白也有明显升高，而其mRNA表达水平却没有变化，提示CYP3A1和CYP3A2的诱导机制可能不同。

RESULTS: Image analysis of two-dimensional map revealed 72 differentially expressed proteins between MHCC97-H and MHCC97-L cells, and 9 protein peaks (including M2 pyruvate kinase,α-subunit of ATP synthase, heat shock protein 60, Toll-like receptor 9, flavoprotein oxygenase, pro-calreticulin, manganese superoxide dismutase, nm23-H1, and G-protein-coupled receptor kinase 5) were further identified by tryptic digestion, peptide mass fingerprinting and mass spectrometry.

结果： 2D图谱显示共有72个差异蛋白条带，共鉴定出了9个差异蛋白，分别为M2型丙酮酸激酶、ATP合成酶α亚单位、热休克蛋白60、Toll样受体9、含黄素单加氧酶、钙网硬蛋白前体、锰超氧化物岐化酶、nm23-H1、G-蛋白偶连受体激酶5；其中4个蛋白在高转移细胞株MHCC97-H中表达升高， 5个蛋白在低转移细胞株MHCC97-L表达升高。

They were named respectively as muramidase-released protein; succinate dehydrogenase/fumarate reductase flavoprotein subunit (SDHFp/FRDFP, extracellular); bacterial trigger factor; elongation factor G (EF-G, extracellular); PTS system sorbose subfamily I (PTS/MAN-ⅡAB, cytoplasmic); pyruvate kinase; phosphofructokinase, PK.

经MALDI-TOF-MS鉴定，这8个蛋白分别为溶菌酶释放蛋白、琥珀脱氢酶／延胡索酸还原酶黄素蛋白亚单位(SDHFp/FRDFp)、触发因子、延长因子G、细菌糖磷酸转移酶系统甘露糖特异ⅡAB亚单位、丙酮酸激酶、6-磷酸果糖激酶(6-PFK)、色氨酸－tRNA合成酶。

A diverse range of effects:on protein solubility and secretion was observed,and these effects were highly dependent on the particular chaperone.In the host strain over-expressing chaperone SecB harboroing pSecB,the amount of total periplasmic proteins was increased by about 71％and the activities of alkaline phosphatseand GL-7-ACA acylase(GL7A)were increased by about 54％and l.5-fold respectively.In the host strain over-expressing chaperone GroEL bearing pGroEL,the amount of total periplasmic proteins was increased by 52％,and the activity of penicillin G acylasewas in creased by about 76％;about 90％of Hexameric Calcitonin(Cal6)inclusion body and l5％of MS2interleukin3(MS2-HIL3)inclusion body became soluble respectively.All the data were compared with the parental strains un-bearing plasmdis pSecB or pGroEL.

在过量表达SecB的宿主菌中，周质空间分泌蛋白总量较对照组提高了约71％，GL-7-ACA酰化酶在周质空间酶的活力较对照组提高了约1.5倍，碱性磷酸酯酶在周质空间酶的活力较对照组提高了约54％；在过量表达GroEL的宿主菌中，周质分泌蛋白总量较对照组提高了约52％，青霉素G酰化在周质空间酶的活力较对照组提高了约76％，鲑鱼降钙素六聚体的可落性组分的比例由原来的45％增加到约90％，而MS2-人白介素-3融合蛋白的包涵体有约15％转变为可溶性组份。

The result of immobilization showed that hydrophile membranes were more advantageous than hydrophobe ones to act as immobilization carrier. Crosslinking organophosphorus hydrolase by glutaraldehyde on polyethersulfone membrane is a simple and practical immobilization method. An appropriate amount of bovine serum albumin and crosslinker is necessary to get a good result of immobilization. In the biodegradation of methyl-parathion through enzyme membrane reactor, the effect of immobilized enzyme amount, flow rate of peristaltic pump, pH of the feed, and the methyl-parathion concentration on biodegradation rate was studied. It was shown that the biodegradation rate increased with immobilized enzyme amount and substrate concentration. Biodegradation rate didn't increase when the folw rate of the peristaltic pump was larger than 7 ml/min.

然后，研究固定化酶降解甲基对硫磷，从不同材料的膜载体固定有机磷水解酶的比较中，发现亲水膜比疏水膜适合作固定化酶载体；在此基础上，将有机磷水解酶固定在聚醚砜微滤膜上，并制成酶膜生物反应器，用于降解甲基对硫磷；戊二醛化学交联法固定时，酶液与10%牛血清白蛋白溶液的比例在3：1~2：1，与交联剂的比例在7：2~7：4范围内，固定化效果较好，并发现牛血清白蛋白和膜载体有降低酶活损失的作用；将固定化的酶膜装于酶膜反应器降解甲基对硫磷，实验结果表明：反应器的降解速率与固定化酶量呈正比，也随底物浓度的增加而加快，当流速低时，降解速率随流速增大而加快，到7ml/min以上时，降解速率不随流速脑黾佣洌到獾淖畹团ǘ任？

Nineteen spots were changed significantly after FA treatment.Thirteen proteins were identified by peptide mass fingerprinting or peptide sequence analysis,including putative nucleoside diphosphate kinase,elongation factor 1-gamma,triosephosphate isomerase,60S acidic ribosomal protein P0,heat shock protein 75 kDa,similar to heat shock 70kD protein binding protein,annexin I,hypothetical protein FLJ34423,microtubule-actin crosslinking factor 1,lamin B2,ATP synthase alpha chain,mitochondrial precursor,proteasome subunit alpha type 6.These identified proteins involved in energy metabolism,translation and RNA processing,protein folding,redox regulation,cell structure and cell signaling.

双向凝胶电泳结果显示，甲醛刺激后19个蛋白斑点发生变化，肽指纹图谱及肽序列标签鉴定了其中13个蛋白斑点，已鉴定的蛋白包括二磷酸核苷酸激酶、延长因子1-γ，磷酸丙糖异构酶、60S酸性核糖体蛋白P0、75kDa热休克蛋白、70kD热休克蛋白样结合蛋白、钙依靠磷脂结合蛋白I、假想蛋白FLJ34423、微管-肌动蛋白交叉连接因子1、核纤层蛋白B2、ATP合成酶α链、蛋白酶体α亚基6，这些蛋白功能涉及转录调节、蛋白折叠、信号传导、能量代谢、细胞骨架等各个方面。

Thirteen proteins were identified by peptide mass fingerprinting or peptide sequence analysis,including putative nucleoside diphosphate kinase,elongation factor 1-gamma,triosephosphate isomerase,60S acidic ribosomal protein P0,heat shock protein 75 kDa,similar to heat shock 70kD protein binding protein,annexin I,hypothetical protein FLJ34423,microtubule-actin crosslinking factor 1,lamin B2,ATP synthase alpha chain,mitochondrial precursor,proteasome subunit alpha type 6.These identified proteins involved in energy metabolism,translation and RNA processing,protein folding,redox regulation,cell structure and cell signaling.

双向凝胶电泳结果显示，甲醛刺激后19个蛋白斑点发生变化，肽指纹图谱及肽序列标签鉴定了其中13个蛋白斑点，已鉴定的蛋白包括二磷酸核苷酸激酶、延长因子1-γ，磷酸丙糖异构酶、60S酸性核糖体蛋白P0、75kDa热休克蛋白、70kD热休克蛋白样结合蛋白、钙依赖磷脂结合蛋白I、假想蛋白FLJ34423、微管-肌动蛋白交叉连接因子1、核纤层蛋白B2、ATP合成酶α链、蛋白酶体α亚基6，这些蛋白功能涉及转录调节、蛋白折叠、信号传导、能量代谢、细胞骨架等各个方面。

BST16 was also proved to encode gene of PSII lOkD protein by protein prosite and conserve domain analysis. The protein encoded by BST16 contained a conserve domain PsbR of PSII lOkD protein from 48 to 140 and its carboxy terminal had an ADH_SHORT prosite of Short-chain dehydrogenases/reductases family signature from 94 to 122, which showed the protein encoded by BST16 would be a reductases.

对该基因可读框架编码的蛋白进行功能位点和结构功能域的分析，同样证明了该基因为PSⅡ 10kD蛋白编码基因，其氨基酸摘要序列 48刁位含有一个保守的光合系统＊ 10kD蛋白结构域 PSbR，在蛋白的猿基端有一个保守的短链脱氢酶／还原酶家族特征序列ADHSHORT（Short－chain dehydrogenases／reductases family signature），位于94－122位，说明了该蛋白可能具有还原酶活性。

Five expressed genes contributed to the flowering process and embryonic development such as prolamine, gene for allergenic protein, chalcone synthase, putative peptide methionine sulfoxide reductase and acyl carrier protein Ⅱ gene were chosen to hybridize with a 1510-RNA-blot (time-point phenotypes) array. The 1510 RNA blots were prepared into an RNA array for the purpose of conferring and validating global transcriptional profiles on booting, flowering and filling quantitatively and qualitatively at a statistic level. The results verified: Prolamine and allergenic protein were high-expressed in filling grains after being regulated by development, Chalcone synthase and methionine sulfoxide reductase were high-expressed in the leavies induced by light, in the root stressed for nitrogen deficiency.

为了全局性分析基因表达谱以及进一步验证它们与开花过程的相关性，选取参与水稻开花过程和胚胎发育过程的基因：查尔酮合酶、酰基载体蛋白Ⅱ、醇溶蛋白、S-腺苷基甲硫氨酸还原酶、过敏反应蛋白等为探针，与1500个水稻RNA斑点阵列进行杂交时，结果证实：醇溶蛋白和过敏反应蛋白受发育调节在乳熟成穗中高表达，查尔酮合酶和S-腺苷基甲硫氨酸还原酶在叶片中受光诱导、在根中受缺氮胁迫高表达。

Do you know, i need you to come back

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Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。