重组人生长激素降解产物的检测分析

本文详细介绍了Ｓ－Ｂ电导率模型确定泥质砂岩储层含水饱和度的基本原理以及相应参数的确定方法。用该方法处理了大庆地区的树１３１井、萨南５丁４－检１３２井及杏９丁３检１３４井的一些层位，并将求得的含水饱和度与岩心分析的含水饱和度进行了对比，吻合得比较好。这说明了应用Ｓ－Ｂ模型确定储层的含水饱和度是可行的，结果比较好。     

质粒pGH/BL_(21)在大肠杆菌中表达条件的研究

目的　寻找质粒 pGH/BL2 1在大肠杆菌中的最佳表达条件。方法　当它在大肠杆菌中表达时 ,考查不同IPTG诱导浓度及不同的诱导时间对表达产物的影响 ,表达产物的量以SDS -PAGE电泳方法分析。结果　该基因的最佳表达条件为 :IPTG诱导浓度 1.0mmol/L ,诱导时间 4h。结论　IPTG对不同的基因在不同的表达体系中需不同的诱导条件 ,只有找到它的最佳诱导条件才能保证终产物的最大产率     

胃癌组织中核基质蛋白的变化

目的：研究胃癌组织核基质蛋白的改变。方法：应用SDS－PAGE技术及Geneools定量分析软件，对22例胃癌组织及正常组织的核心基质蛋白进行了研究，结果：胃癌组织与正常组织比较，Mr为30000，28000的核基质蛋白表达量明显减少（P＜0．05），不同分化类型及不同临床分期胃癌组织间比较，此种核基质蛋白表达量差异无统计学意义（P＞0．05），结论：胃癌组织中核基质蛋白的改变可能在肿瘤的早期已经发生，是胃癌发生的早期分子事件。     

幽门螺旋菌菌体蛋白质初步研究

用聚丙烯酰胺凝胶电泳(SDS—PAGE)分析幽门螺旋菌(HP),其超声粉碎和冻融抗原有7条主要蛋白质条带,分子量为71kd、62kd、58kd、51kd、29kd、22kd和12kd。免疫印迹法发现HP100kd免疫性强、特异性高。HP外膜蛋白质分子量主要为71kd、58kd、29kd和12kd,HP不同林之间SDS—PAGE图谱相似,但与空肠弯曲菌和结肠弯曲菌明显不同。超声粉碎和冻融处理HP后主要蛋白质条带相同,冻融抗原制备简单,适宜作为ELISA使用抗原。     

Proteolytic artifacts in SDS-PAGE analysis of selected periodontal pathogen:

The aim of the study was to examine whether proteolytic artifacts, which result in a loss and poor resolution of protein bands, occur during sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis analysis of cellular proteins from selected proteolytic ( Porphyromonas gingivalis, Prevotella nigrescens and Treponema denticola ) and non-proteolytic ( Fusobacterium nudeatum ) bacteria. Conditions to limit or prevent proteolysis were also investigated. Bacterial cells were incubated in solubilizing buffer (SDS +β mercaptoethanol) at room temperature for various periods of time before boiling. A control assay consisted of trichloroacetic acid-treated bacterial cells. Cellular proteins were separated by electrophoresis and stained with Coomassie blue. Proteolysis occurred very rapidly in the case of P. gingivalis (<30 s), whereas a longer incubation time (>1 h) was required to observe similar effects in P. nigrescens and T. denticola. No proteolysis was observed for F. nudeatum. In all cases, heat (100°C) and low pH (<4) treatments of bacterial cells could avoid production of proteolytic artifacts. Incorporation of specific protease inhibitors before solubilization of bacteria could also prevent proteolysis. More particularly, N -α- p -tosyl- l -lysine chloromethyl ketone (TLCK), iodoacetamide and diisopropylfluorophosphate (50 mM) were highly efficient for P. gingivalis, P. nigrescens and T. denticola , respectively. When outer membranes of P. gingivalis were prepared in the presence of TLCK, numerous additional protein bands, not seen in the absence of TLCK, were detected. The present study suggests that specific protease inhibitors, effective in preventing proteolysis. should be identified and added during cell fractionation and protein purification procedures.     

球形幽门螺杆菌生物学性状及蛋白产物研究球形Hp系列研究之二

目的：研究球形幽门螺杆菌生物学性状和蛋白产物发生的改变。方法：通过延长培养时间和亚抑菌浓度抗生素－－灭滴灵的作用，分别获得两类球形幽门螺杆菌（Ｈｐ）。结果：两种不利于Ｈｐ生长的环境在引起Ｈｐ发生球变的同时，每毫升菌落形成单位（ＣＦＵ／ｍｌ）减少，尿素酶活性降低。结论：球形Ｈｐ尿素酶活性低下，且在常规培养基上不能生长。抗生素的作用比延期培养更快使Ｈｐ发生球形变异。聚丙烯酰胺凝胶电泳蛋白质条带分析发现抗生素诱变的球形Ｈｐ与正常形态Ｈｐ基本相似，与延期培养的Ｈｐ明显不同。     

Characterization of a monoclonal anti-Bw4 antibody (Tü109): Evidence for similar epitopes on the Bw4 and Bw6 antigens

The production and serologic, as well as immunochemical properties of a cytotoxic murine IgG monoclonal antibody (Tü109) that precipitates HLA-class I molecules, are described. In the microcytotoxicity assay Tü109 supernatant was demonstrated on a panel of 424 HLA-ABC, -DR, -DQ, -MT typed normal Caucasian blood donors to define an epitope on HLA-B locus molecules in great association with the supertypic specificity Bw4. Reactivity of supernatant showed MHC linked inheritance of the Tü109 determinant and discriminated the HLA-Bw4/Bw6 associated HLA-B locus split antigens. Weak or lack of binding on lymphocytes from some HLA-Bw4 heterozygous individuals, particularly typing for HLA-Bw44, appeared to be due to qualitative and/or quantitative variations of HLA-B locus molecules on the cell surface. With Tü109 ascites fluid, however, extra-reactivity on all HLA-Bw6⁺ cells was demonstrated. Preferential binding of supernatant to HLA-Bw4, but reactivity of ascites fluid with HLA-Bw6⁺ molecules in addition, was furthermore confirmed by IEF analysis of antigens immunoprecipitated with Tü109 from cell lysates. Thus the antibody may help to analyze the evolutionary relationship of the diallelic specificities Bw4 and Bw6.     

Mouse monoclonal antibodies to the human C3b receptor

Mouse monoclonal antibodies were raised against the human C3b receptor (CR1) molecule that had been purified from solubilized erythrocytes membranes. Four hybridomas were selected, cloned and expanded because their supernatants reacted strongly with insolubilized CR1 by ELISA and intensely stained B-dependent areas of the spleen and glomerular podocytes by indirect immunofluorescence. The four monoclonal antibodies, named J3D3, J8B10, J3B11 and J7C2, were IgG1 immunoglobulins. J3D3 immunoprecipitated two protein bands of apparent mol. wts 200,000 and 220,000 from 125I-surface-labeled human erythrocytes, which correspond to the two major allotypic forms of CR1. By indirect immunofluorescence, monoclonal antibodies stained polymorphonuclear leucocytes (PMN), most peripheral blood B-cells and a small subset of peripheral blood T-cells. J3D3 bound to CR1 on erythrocytes, PMN and lymphocytes with an affinity of 1-3 X 10(9) M-1 and recognized 170-1330 antigenic CR1 sites with an average of 740 sites/erythrocyte in 100 healthy individuals, approx. 50,000 sites/PMN and 15,000 sites/lymphocyte. There was a bimodal distribution of CR1 numbers on erythrocyte in the normal population. The four monoclonal antibodies similarly inhibited CR1-mediated decay of preformed cell-bound alternative- and classical-pathway C3 convertase sites. Two antibodies, J3D3 and J3B11, inhibited C3b-dependent rosette formation with lymphocytes, although much less efficiently than F(ab')2 polyclonal anti-CR1 antibody. Differences that were observed in the relative capacity of the antibodies to inhibit some of the functions of CR1 and in their ability to compete for binding of 125I-J3D3 to CR1 on erythrocytes, suggested that they are directed against different epitopes on CR1. Monoclonal antibodies provide useful means to assess and analyze the biological and immunoregulatory functions of the C3b receptor.     

Structural difference between the two forms of guinea-pig beta 2-microglobulin and their occurrence in inbred guinea-pig strains

The structural difference between two forms (basic and acidic) of guinea-pig beta 2-microglobulin (beta 2m) has been established. Both forms are present in urine from inbred guinea-pig strains. The beta 2m forms were each digested with carboxypeptidase Y and carboxypeptidase A contaminated with carboxypeptidase B. Released amino acids were separated from remaining protein, dansylated and analysed by 2-dimensional TLC on polyamide layer sheets. From the results it was concluded that the basic beta 2m form has lysine and the acidic beta 2m form has asparagine as their respective C-terminal amino acids. The acidic form is also 1 amino acid (lysine) shorter than the basic form, which is supported by electrophoretic studies on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The presence of the 2 forms of beta 2m in urine from inbred guinea-pig strains 2 and 13, shown by gel filtration and ion exchange chromatography, makes it unlikely that the 2 forms are a result of genetic polymorphism.