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41.
Generation of a recombinant Moloney murine leukemia virus carrying the v-src gene of avian sarcoma virus: transformation in vitro and pathogenesis in vivo. 总被引:6,自引:5,他引:1 下载免费PDF全文
A Moloney murine leukemia virus (M-MuLV) recombinant carrying the v-src gene of avian sarcoma virus was generated by the introduction of a cloned portion of v-src from Schmidt-Ruppin A avian sarcoma virus into a molecular clone of M-MuLV provirus at the recombinant DNA level. The v-src sequences (lacking a portion of the 5' end of v-src) were inserted into the p30 region of the M-MulV gag gene so that M-MuLV gag and v-src were in the same reading frame. Transfection of this chimeric clone, pMLV(src), into NIH 3T3 cells which were constitutively producing M-MuLV gag and pol protein resulted in the formation of foci of transformed cells. Infectious and transforming virus could be recovered from the transformed cells. This virus was designated M-MuLV(src). M-MuLV(src)-transformed cells contained two novel proteins of 78 and 90 kilodaltons. The 78-kilodalton protein, p78gag-src, contained both gag and src determinants, exhibited kinase activity in an immune kinase assay, and is probably a fusion of Pr65gag and src. The 90-kilodalton protein, which is of the appropriate size to be the gPr80gag fused to src, contained gag determinants as well as a V8 protease cleavage fragment typical of the carboxy terminus of avian sarcoma virus pp60src. However, it could not be immunoprecipitated with an anti-v-src serum. M-MuLV(src)-transformed cells showed elevated levels of intracellular phosphotyrosine in proteins, although the elevation was intermediate compared with cells transformed with wild-type v-src. M-MuLV and amphotropic murine leukemia virus pseudotypes of M-MuLV(src) were inoculated into newborn NIH Swiss mice. Inoculated mice developed solid tumors at the site of inoculation after 3 to 6 weeks, with most animals dying by 14 weeks. Histopathological analysis indicated that the solid tumors were mesenchymally derived fibrosarcomas that were both invasive and metastatic. 相似文献
42.
Molecular hydrogen inhibits nitrogenase activity in Anabaena pre-illuminated with red or blue light. The inhibitory effect of molecular hydrogen decreased in the presence of oxygen and several electron acceptors. When NH4Cl and urea were added simultaneously with molecular hydrogen, marked synergistic inhibitory effects took place. The inhibitory effect of molecular hydrogen disappeared or was weakened after the suppression of hydrogenase activity. The addition of O2 and electron acceptors to systems showed no enhancing effect on the C2H2-reducing activity. 相似文献
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44.
甲基苄基亚硝胺对与人、大鼠和鸡肝脏或食管上皮细胞混合培养的V_(79)细胞的致突变作用 总被引:3,自引:0,他引:3
用人,雄性Wistar大鼠和雄性莱亨鸡的肝细胞,或食管上皮细胞与V_(79)细胞混合培养,观察甲基苄基亚硝胺(MBN)经肝细胞或食管上皮细胞代谢后,能否产生活性代谢产物,从而诱导V_(79)细胞突变及其它有关的生物学反应。研究结果初步表明,大鼠和鸡肝细胞可以明显代谢MBN产生活性代谢产物,诱导混合培养的V_(79)细胞SCE和微核增高以及6-TG抗性突变。人胎儿肝细胞也可以代谢这种亚硝胺并诱导V_(79)细胞6-TG抗性突变,但对SCE和微核诱导明显偏低。说明这三种肝细胞代谢MBN在性质上是比较近似,可能在代谢激活程度上有一定差异。在食管上皮细胞和V_(79)细胞混合培养的实验中,大鼠食管上皮可以明显代谢MBN诱导V_(79)细胞SCE和微核的增高及6-TG抗性突变。鸡食管上皮细胞未见有明显的代谢MBN引起V_(79)细胞突变的作用。26只雄性莱亨鸡喂以MBN实验最长者达975天,总剂量为1674毫克,结果未见一例鸡发生食管癌。结果表明鸡的食管上皮可能对某些挥发性亚硝胺的致癌作用是不敏感的,目前没有证据支持在林县高发区中所发现的鸡咽食管癌是由于已找到的挥发性亚硝胺引起的。人食管上皮细胞具有一定的代谢MBN产生有致突变作用的代谢产物的能力,然而人与人之间有明显的个体差异。和大鼠相比,人食管代谢MBN诱导V_(79)细胞突变的能力明显较低,有关亚硝胺在高发区人食管癌发生中的作用还有待进一步深入研究。 相似文献
45.
人类活动影响下新疆生态环境的一些变化 总被引:4,自引:0,他引:4
人类活动引起的新疆生态环境变化主要有以下几方面:1.河流流程缩短,河水矿化度和泥沙含量增加;2.湖泊水位下降,湖面缩小或变干,湖水矿化度升高;3.森林破坏,面积缩小;4.草场的产草量降低,牧草质量下降; 5.土壤次生盐渍化、沼泽化和肥力下降;6.土地沙漠化不断扩大。 相似文献
46.
用含80%1,4-丁二醇的混合溶剂,以胰蛋白酶酶促,由去八肽胰岛素(DOI)合成了去六肽胰岛素(DHI),总产率为35%。1,4-丁二醇的溶解性能好,在浓度高达80—90%时不明显抑制酶活力,DOI的氨基无需保护,溶液中无高聚物或沉淀形成。 相似文献
47.
用人肺鳞癌细胞LTEP-78细胞系免疫Blab/c小鼠获得3株抗人肺癌细胞的单克隆抗体杂交瘤系。其中BLTI-01株经六次克隆化培养,体外传代8个月以上。BLTI-01与白细胞抗原及血型抗原基本上无交叉反应;与骨髓细胞无交叉反应;与癌胚抗原和胎甲球蛋白不相关;与肺鳞癌、肺腺癌细胞系及部分其它肿瘤细胞呈阳性反应。 相似文献
48.
49.
Alloimmune murine thymus-derived cytotoxic lymphocytes (CTL) generated in vivo or in vitro are shown to lyse antigen-nonspecific target cells (tumor cells, Con A, and LPS blasts) following treatment of CTL with an oxidizing agent, sodium periodate (NaIO4). It has been shown that NaIO4 oxidizes terminal sialic acid residues of cell surface macromolecules. The presence of reactive aldehyde groups, generated by NaIO4 modification, is required for the expression of antigen-nonspecific cytotoxicity because treatment of modified cells with a reducing agent such as potassium borohydride (KBH4) resulted in the abrogation of cytotoxicity. However, KBH4 treatment of unmodified or NaIO4-modified CTL has no effect on antigen-specific cytotoxicity. The modification of CTL by NaIO4 is sufficient to lead to the formation of lymphocyte-target cell conjugates and lysis of bound targets. Monoclonal antibodies directed against the Lyt-2 antigens of CTL, but not Lyt-1 antigens, in the absence of complement inhibited the nonspecific cytotoxicity resulting from NaIO4 modification of effector lymphocytes. These findings suggest that the mere interaction with or perturbation of appropriate cell surface molecule(s) of effector lymphocytes such as Lyt antigens by receptor-ligand interaction in SCMC or by NaIO4 modification in ODCC may lead to the expression of cytotoxicity. The present studies demonstrate a functional role of surface carbohydrates on CTL in cell-to-cell recognition and interactions. Furthermore, the results suggest that target cell modification is not a requisite for recognition and lysis in an antigen-nonspecific cytotoxic system such as ODCC. However, partial blocking of ODCC by alloantibodies directed against the H-2 of unmodified target cells suggests that NaIO4-modified CTL recognize unrelated target H-2 antigens. The implication of these findings on the molecular mechanism of cell-mediated cytotoxicity is discussed. 相似文献
50.
An xenogeneic rat anti-mouse T-cell serum, designated RAT*, has been shown to block the cytolytic activity of cytotoxic T lymphocytes (CTL) at a postbinding step. RAT* serum or the IgG fraction was extensively absorbed with the target cell, P815, a DBA mastocytoma, and used with or without further absorption to immunoprecipitate specific molecules from radiolabeled membrane extracts of CTL derived from either in vivo-allosensitized mice or from cytotoxic clones maintained in in vitro cultures. Cell surface sialic acid residues were labeled by oxidation with sodium periodate (NaIO4) and reduction with tritiated sodium borohydride ([3H]NaBH4). Alternatively, cell surface proteins were labeled with 125I by lactoperoxidase-catalyzed iodination. Nonidet P-40 (NP-40)-solubilized radiolabeled membranes were then immunoprecipitated with RAT* serum and analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). Three membrane-associated molecules of 95,000, 140,000 and 180,000 Mr were found by such analysis. The sensitivity of these three molecules to trypsinization and their susceptibility to labeling with [3H]NaBH4 suggested that they are glycoproteins. Moreover, when RAT* serum or the IgG fraction was absorbed with various cell types, its ability to immunoprecipitate the three molecules correlated with its ability to block cytolysis. Adsorption of RAT* serum with CTL, but not with nonimmune thymocytes, significantly reduced the ability of RAT* serum to inhibit cytotoxicity and to immunoprecipitate the 95k, 140k, and 180k molecules. Thus, these findings suggest that one or more of these cell surface molecules of CTL may be involved in the cytolytic process. 相似文献