His-tag不影响RSV重组蛋白G1F/M2的免疫原性

目的：观察His-tag是否影响RSV重组蛋白G1F/M2的免疫原性。方法：PCR扩增G1和F/M2基因片段,插入表达载体pET-His和pET-DsbA-His中,转化E.coli BL21（DE3）,IPTG诱导表达,采用Ni^＋螯合亲和层析法纯化得His-G1F/M2和DsbA-His-G1F/M2,将后者用凝血酶消化,再经Ni＋螯合亲和层析法纯化得G1F/M2,将His-G1F/M2和G1F/M2免疫BALB/c小鼠,用ELISA测定抗体滴度,MTT法测定细胞毒性T细胞活性（CTL）。结果：两种蛋白在BALB/c小鼠中诱导的RSV特异性抗体和CTL活性无显著差异。结论：His-tag不影响RSV重组蛋白G1F/M2的免疫原性。     

Isolation of Trypanosoma brucei variant specific antigen mRNA by immunoprecipitation of polysomes

The mRNA encoding the variant specific antigen of Trypanosoma brucei has been prepared by immunoprecipitation of polysomes. Polysomes carrying the variant specific antigen account for approx. 3% of the total polysomes. The mRNA thus produced is active in the mRNA dependent rabbit reticulocyte lysate in vitro protein synthesis system and directs the synthesis of a polypeptide of 60 000 daltons which co-migrates both with ¹²⁵I-labelled purified variant specific antigen and with antigen immunoprecipitated from reticulocyte lysate charged with total polyadenylated mRNA from the same clone. The mRNA is being used both to prepare cDNA clones and to prepare high specific radioactivity cDNA to be used to screen a gene bank for clones containing variant specific antigen coding sequences.     

软组织平滑肌肉瘤中p16基因的甲基化检测

目的 探讨软组织平滑肌肉瘤（ＬＭＳ）中p16基因ＩＮＫ４Ａ的甲基化状态及其与p16表达的关系。方法 应用ＭＳＰ法检测３８例软组织平滑肌肉瘤,１０例平滑肌瘤及５例正常平滑肌组织中p16基因ＩＮＫ４Ａ的甲基化状态,用免疫组织化学ＳＰ方法检测p16蛋白表达情况。结果 ３８例ＬＭＳ中９例发生异常甲基化,异常甲基化率为２３.7%（９／３８）。其中,７例p16蛋白表达阴性,２例p16蛋白弱阳性,在p16蛋白表达阴性的ＬＭＳ中,异常甲基化率为５０％（７／１４）。结论 p16基因第一外显子启动子区５‘CpG岛的异常甲基化是导致p16基因失活、蛋白缺如的重要基因外机制,并可能参与肿瘤的发生。     

Multiple Copies of the Upstream Regulatory Region of the Major Capsid Protein Gene of Bacteriophage MB78 Inhibit Phage Morphogenesis

The 2.311 kb EcoRI F fragment of bacteriophage MB78 has been cloned in multicopy vectors pUC19 and pCR90. Salmonella typhimurium strains carrying such plasmids cannot support development of phage MB78 while other Salmonella phages like P22 and 9NA grow normally. Most of the phage MB78 induced functions are normal in such transformed hosts but proper maturation of the phage particles does not take place. Deletion of 138 bp from the 3 end of the cloned fragment reverses the inhibitory effect. Analysis of nucleotide and the deduced amino acid sequence of a 1.2 kb HindIII-SalI fragment of the phage genome which overlaps the 138 bp confirms that this part contains the upstream regulatory region of the major structural protein gene. It seems that in presence of multiple copies of the upstream regulatory region (which includes a number of promoter like sequence) of the coat protein gene, the maturase gene is down regulated and this is effective only in cis, a situation quite similar to that of Q RNA phages.     

Role of Apolipoprotein A-I in Steroid-Induced Activation of DNA and Protein Synthesis in Hepatocytes

It is demonstrated that anabolic effect of steroid hormones is produced by steroid-apolipoprotein A-I complexes. These complexes accelerate not only protein, but also DNA synthesis and produce both cell hypertrophy and hyperplasia. For realization of their anabolic effect steroid hormones are modified by - and -reductases in resident macrophages.     

Preferential presentation of herpes simplex virus T-cell antigen by HLA DQA1*0501/DQB1*0201 in comparison to HLA DQA1*0201/DQB1*0201

The HLA DQA1 locus is polymorphic. Haplotypes containing HLA DQA1*0501, but not HLA DQA1*0201, together with HLA DQB1*0201 are associated with Grave's disease and celiac sprue. In this report, we demonstrate a functional correlate of DQA1 polymorphism. T cells infiltrating a herpes simplex virus (HSV) lesion from a HLA DQ 2,7 individual yielded a virusspecific CD4+ clone restricted by DQ2. Presentation of viral peptide and protein segregated with DQA1 allele, because cell lines bearing DQA1*0501/DQB1*0201 heterodimers presented antigen in proliferation and cytotoxicity assays much more efficiently than cell lines bearing DQA1*0201/DQB1*0201. Binding of viral peptide to cell lines bearing DQA1*0201, in comparison to DQA1*0501, was only moderately reduced and may not explain this effect. Truncation and substitution analyses of peptide binding and T-cell activation were performed to determine which viral peptide residues contacting TCR might therefore be presented in an altered conformation by DQA1*0201/DQB1*0201. Residues 432, 435, 437, 438, and 440 (position P1, P4, P6, P7, and P9) contributed to DQ2 binding, whereas residues 431, 433, 434, and 436 (positions P-1, P2, P3, and P5) contributed to TCR contact. Differential presentation of peptide by HLA DQ2 heterodimers varying at the DQA1 locus may have relevance to host defense and the pathogenesis of HLA DQ2-associated autoimmune diseases. Human Immunology 53, 195-205 (1997).     

Identification of a Strong Promoter of Bacteriophage MB78 that Lacks Consensus Sequence Around Minus 35 Region and Interacts with Phage Specific Factor

A strong promoter of bacteriophage MB78 does not have minus 35 consensus sequence although it has a TGn motif immediately upstream of minus 10 sequence as well as the AT rich UP element. It is efficiently recognised by the sigma 70 RNA polymerase, however, a phage-specific factor competes with sigma 70 RNA polymerase for binding to this region, the binding of the factor being stronger than that of the polymerase. Contrary to the reports in the literature the polymerase appears not to bind to the UP element whereas the phage-specific factor does. The latter seems to be involved in the regulation of the promoter activity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of o,p-dichlorodiphenyldichloroethane and perthane in vitro on glutathione reductase activity in the adrenals of dogs and guinea pigs

o,p-Dichlorodiphenyldichloroethane (o,p-DDD) and Perthane, when added in a concentration of 312 M to homogenate and cytoplasmic fraction of dog adrenals, activate glutathione reductase. In a concentration of 156 M, o,p-DDD and Perthane do not affect glutathione reductase activity of the dog adrenals. When given in vitro, o,p-DDD and Perthane activate glutathione reductase of the guinea pig adrenals. o,p-DDD has no effect on glutathione reductase activity of the cytoplasmic fraction of dog liver and kidney, thus confirming the high specificity of its effect on the adrenal cortex.Laboratory of Pathophysiology, Institute of Endocrinology and Metabolism, Kiev. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 2, pp. 159–161, February, 1978.     

Cardiac ankyrin repeat protein is preferentially induced in atrophic myofibers of congenital myopathy and spinal muscular atrophy

Cardiac ankyrin repeat protein (CARP), which is structurally characterized by the presence of four ankyrin repeat motifs in its central region, is believed to be localized in the nucleus and to participate in the regulation of cardiac-specific gene expression in cardiomyocytes. However, we recently found that CARP was induced in skeletal muscle by denervation, leading us to speculate that CARP may be induced under some pathological conditions. In the present study, we immunohistochemically analyzed the expression of CARP in 11 cases of spinal muscular atrophy (SMA) and 14 cases of congenital myopathy. In SMA, CARP was expressed selectively in severely atrophic myofibers, suggesting that CARP expression may reflect the status of muscle atrophy. Furthermore, in the congenital myopathies, the expression patterns of CARP were distinct among the subtypes, which included nemaline myopathy, myotubular myopathy, central core disease, and congenital fiber type disproportion. Although CARP was preferentially expressed in severely damaged myofibers in nemaline myopathy, it was not detected in central core disease. These findings suggest that immunohistochemical evaluation of CARP may be helpful in the diagnosis of SMA and the congenital myopathies.