分子克隆和表达人类的肿瘤坏死因子与小鼠肿瘤坏死因子和比较。
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Marmenout, Fransen L, Tavernier J, Van der Heyden J, Tizard R,川岛E,肖,约翰逊MJ,义符D,穆勒R, et al。
分子克隆和表达人类的肿瘤坏死因子与小鼠肿瘤坏死因子和比较。
欧元。1985 11月4日,152(3):515 - 22所示。
- PubMed ID
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3932069 (在PubMed]
- 文摘
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u - 937细胞,单核细胞的线源自人类组织细胞的淋巴瘤,是人类肿瘤坏死因子(TNF)诱导分泌到培养基中,用于肿瘤坏死因子mRNA的准备。生物活性的量化在非洲爪蟾蜍光滑的卵母细胞注射系统。肿瘤坏死因子mRNA由梯度离心浓缩,这size-fractionated信使rna被用于合成cDNA和插入到独特的pAT153 PstI网站。重组质粒含有人类TNF cDNA被菌落杂交选择使用一个内部碎片鼠标TNF cDNA克隆(Fransen L。穆勒,R。Marmenout,。Tavernier, J。Van der Heyden, J。川岛,E。Chollet,。Tizard, R。范Heuverswyn, H。Van Vliet,。Ruysschaert, m . r . &菲尔w(1985)核酸研究》13日4417 - 4429)作为探针。这个人类TNF cDNA序列是同意由Pennica et al。(Pennica D。Nedwin, g . E。海弗利克,j·S。,Seeburg, p . H。Derynck, R。, Palladino, M. A., Kohr, W. J., Aggarwal, B. B. & Goeddel, D. V. (1984) Nature (Lond.) 312, 724-729]. The 157-amino-acid-long mature sequence is about 80% homologous to mouse TNF and its hydrophilicity plot is also very similar, in spite of the apparent species specificity of TNF. In contrast to mouse TNF, it contains no potential N-glycosylation site. When compared to other cytokines, like IFN-beta, IFN-gamma, or IL-2, there is a remarkably high preference for G X C pairs in the third-letter positions. Expression of the TNF cDNA in monkey COS cells or in Escherichia coli gives rise to a protein having similar biological and serological properties as natural human TNF. A human genomic clone was also identified and sequenced; it was found to be in good agreement with the one recently published by Shirai et al. [Shirai, T., Yamaguchi, H., Ito, H., Todd, C. W. & Wallace, R. B. (1985) Nature (Lond.) 313, 803-806], except for some differences in the introns and 5'-untranslated region.