酶蛋白

The results showed that the total numbers of integral membrane proteins identified based on PAOT,trypsin/chymotrypsin and methanol-modified digestions were 101,59 and 72,which included 67,40 and 31 proteins with GRAVY＞0,respectively.

结果表明，甲醇酶解法鉴定到72个整合膜蛋白，其中平均疏水值大于0的蛋白数为31；胰蛋白酶/胰凝乳蛋白酶酶解法共鉴定到59个整合膜蛋白，其中平均疏水值大于0的蛋白数为40个；PAOT法共鉴定到101个整合膜蛋白，其中平均疏水值大于0的蛋白数为67个。

Recombinant ptotein made up 39.2%(pET-20b-bglA) and 33.5%(pET-28a-bglA) of the total proteins in the intracellular fraction, the solubility proportion of the enzyme up to 32.8%(pET-20b-bglA) and 40.1%(pET-28a-bglA), the activities of the enzyme were 66.8 (pET-20b-bglA) and 71.2 U/mg (pET-28a-bglA). These results showed that E. coli BL21-CodonPlus (DE3)-RIL with argU tRNA, ileY tRNA and leuW tRNA genes helped to improve expression of the enzyme through enhanced identification of the rare codons AGA/AGG, AUA and CUA. Further optimized the conditions for inducing, the solubility proportion of the enzyme was 70.6% at 16 °C and 1.2 times higher than 37 °C. The solubility proportion of the enzyme increased from 12.3% to 32.8% when IPTG concentrations dropped from 1000 μM to 25 μM. The recombinant enzyme was purified by heat treatment, DEAE Sepharose anion-exchange chromatography and TOYOPEARL HW-55F.

maritima MSB8 的-葡萄糖苷酶基因 bglA克隆至表达载体 pET-20b 和 pET-28a，构建重组质粒 pET-20b-bglA 和 pET-28a-bglA，然后转化不同大肠杆菌 E.coli 宿主，Tm-SIGlA 在 E.coli BL21-CodonPlus(DE3)-RIL 中获得高效表达，重组蛋白的表达量为 33.5%(pET-28a-bglA)和 39.2%(pET-20b-bglA)，可溶性比例为 32.8%(pET-20b-bglA)和 40.1%(pET-28a-bglA)，比酶活达 66.8 (pET-20b-bglA)和 71.2 U/mg (pET-28a-bglA)，这些结果表明，E.coli BL21-CodonPlus(DE3)-RIL 宿主带有的 argU tRNA、ileY tRNA 和 leuW tRNA 基因，分别增强对稀有密码子 AGA/AGG、AUA 和 CUA 的识别，有助于提高该酶在 E.coli 中的表达；进一步优化诱导条件，重组 E.coli BL21-CodonPlus(DE3)-RIL/pET-20b-bglA 在 37 ℃下诱导培养，IPTG 浓度由 1000 μM 降至 25 μM，目的蛋白可溶性表达由 12.3%增至 32.8%，提高 1.7 倍，16 ℃下低温诱导实现目的蛋白 70.6%的可溶性表达，较 37 ℃下诱导培养提高 1.2 倍。

The absorption of root system was the main sources of nitrogen in the plant.The effects of matrix ventilation on the abaorption,transportion and metabolism of nitrogen in the root system of cucmber were discussed deeply.The results showed that contents of ammonium,nitrate,free amino acids and soluble protein in cucumber roots were higher in G1 treatment.Contents of ammonium,nitrate and free amino acids in cucumber roots were the trend of the dropping after the first rising in F1 treatment,but contents of soluble protein was lower11.03%than that of the control.

根系吸收是植物体内氮素的主要来源，本研究深入探讨了基质通气状况对黄瓜根系氮素吸收、运输和氮代谢的影响，结果表明G1通气处理根系的铵盐、硝酸盐、游离氨基酸和可溶性蛋白含量均较高；非通气处理(F1处理)根系内铵盐、硝酸盐和游离氨基酸含量呈先升高后降低的趋势，而可溶性蛋白含量始终较低，比对照降低11.03%。G1处理根系硝酸还原酶、亚硝酸还原酶、谷草转氨酶、谷丙转氨酶、谷氨酰胺合成酶活性均较高；而F1处理的酶活性在处理末期显著降低，其中谷氨酰胺合成酶活性比对照降低33.33%。

The effects of jaundice discoloration and lipemia of various concentrations on enzymatic method for determing serum potassium;2. In the experiment the oat protein was extracted from oat flour with ultrasonic-assisted enzymatic method .

采用超声波辅助酶法从燕麦粉中提取燕麦蛋白，研究了料水比、粉碎度、超声时间、超声功率、超声温度以及酶解pH值、酶解时间、酶解温度、加酶量对燕麦蛋白提取率的影响。

There were twelve different hepatocyte proteins interacting with HCBP12 by Yeast two-hybrid, containing human sapien apolipoprotein C-I/B, human sapiens mitochondrion, human sapiens metallothionein 2A, human sapiens beta-2-microglobin,human sapiens carboxylesterase, and human sapiens serpin peptidase inhibitor, with known function and one with unknown function.

利用酵母双杂交技术筛选出12个与HCBP12相互作用的蛋白，其中包括人类载脂蛋白C-I/B、人类线粒体蛋白、人类金属硫蛋白2A、人类β-2微球蛋白、人类羧酸脂酶4和人类丝氨酸肽酶抑制剂等已知功能蛋白及1个未知功能蛋白。

Purified protein samples were separated by 2-DE with 17 cm IPG strips, and the protein spots were detected by fast silver staining. Six protein spots were randomly excised from the silver stained gel for primary analysis by MALDI-TOF MS. Four proteins were identified, and they were 1-aminocyclopropane-1-carboxylate synthase 2, ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit, F11O4.2 and Rubisco small subunit.

根据预实验结果，选用17 cm IPG预制干胶条对纯化的蛋白样品进行2-DE实验，采用快速银染法进行染色，随机挖取银染胶上6个蛋白点进行MALDI-TOF MS分析和数据库检索，鉴定了3个蛋白，它们是：1-氨基环丙烷-1羧酸合酶2，核酮糖-1，5-二磷酸羧化酶/加氧酶大亚基和F11O4.2。

ASES is 59% identical to Artemisia cyclase cDNA clone cASC125, 50% identical to epi-cedrol synthase from A. annua, 48% identical to amorpha-4, 11-diene synthase from A. Annua, 39% identical to the 5-epi-aristolechene synthase from tabacco, 38% identical to vetispiradiene synthase from H. Muticus, 41% identical to the δ-cadinene synthase from cotton. The coding region of cDNA was cloned into procaryotic expression vector pET30a and overexpressed in E. coli BL21 (DE3). The cyclase proteins extracted from bacterial culture were found largely in the insoluble protein fraction. The tissue specific expression patterns were analyzed by RT/PCR.

克隆的倍半萜合酶氨基酸序列与烟草马兜铃烯合酶、莨菪岩兰螺旋二烯合酶、棉花杜松烯合酶的一致性分别为39％，38％和41％；与青蒿柏木脑合酶、紫穗槐二烯合酶和一个推测的倍半萜合酶克隆cASC125的一致性为50％，48％和59％。cDNA编码区序列被克隆进原核表达载体pET-30a，并在大肠杆菌BL21（DE3）中诱导表达，但过量表达的蛋白主要是以不溶性蛋白形式存在。

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003～2007年临床分离的220株Pa，对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况，同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration，MIC)，筛选出对亚胺培南耐药的铜绿假单胞菌，并分析其对其它抗生素的药物敏感率；采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株，采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因，采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因，用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况；同时对产AmpC酶的Pa(25株，含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

Phenylmethylsulfonyl fluoride (174.2 mg/mL), chicken ovomucoid (1000 mg/mL) and soy-bean trypsin inhibitor (1000 mg/mL) could inhibit enzyme activity, which indicated that the enzyme belonged to serine protease group. On plasminogen-free fibrin plates and plasminogen fibrin plates, the fibrinolytic activity had no obvious difference, indicating that the enzyme was a fibrinolytic enzyme which degraded fibrin directly, but not a plasminogen activator which degraded fibrin by activating plasminogen.

此菌株产生的纤溶酶在50℃以下和pH5.0~11.0范围内具有较好的稳定性，最适作用温度为42℃；最适pH值为9.0；Mg2+、Ca2+对此酶有明显的激活作用，而Cu2+能完全抑制酶的活性；174.2 mg/mL的苯甲基磺酰氟、1000 mg/mL的鸡卵类粘蛋白和1000 mg/mL大豆胰蛋白酶抑制剂能完全抑制酶活性，初步说明此酶属于丝氨酸蛋白酶类；体外溶纤作用表明，该酶溶解纤维蛋白的方式是直接溶解，而不是通过激活纤溶酶原。

No immunostaining was seen in RPE. Double staining that Kir2. 1 and glutamine synthetase, Kir2. 1 and glutamine synthetase colocalized well. Intense Kir7. 1 immunolabeling was present on the apical surface of all RPE cells and appeared to extend over the length of the apical processes. Na〓, K〓-ATPase expression varied among RPE cells, but in highly expressing cells, it co-localized with Kir7. 1. Immunoreactivity of Kir2. 1、Kir4. 1 and Kir7. 1 acomplished by ABC immunohistochemistry in monkey and human retinae got the nearly same results as bovine.

间接免疫荧光组织化学显示，Kir2.1蛋白主要分布在Müller细胞与神经元相接触的细胞膜区域；与Müller细胞的特异性标记蛋白质抗谷氨酸合成酶抗体双重标记显示其分布特性重叠，在RPE未检测Kir2.1蛋白的免疫活性存在；Kir4.1蛋白集中分布于Müller细胞的内侧突起，与GS蛋白双重标记显示其分布特性重叠，RPE未检测Kir4.1蛋白的免疫活性存在；Kir7.1主要分布于RPE游离面及其突起的全长，与特异性标记RPE突起的Ezrin蛋白完全重叠，Na〓-K〓ATP酶在不同部位的RPE表达不同，Na〓-K〓ATP高表达的RPE细胞与Kir7.1的分布特性重叠。

Do you know, i need you to come back

你知道吗，我需要你回来

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。