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基因向人类细胞的转移:在GM1神经节苷脂贮积症成纤维细胞中转导噬菌体λplac基因表达

Gene transfer to human cells: transducing phage lambda plac gene expression in GMI-gangliosidosis fibroblasts.

作者信息

Horst J, Kluge F, Beyreuther K, Gerok W

出版信息

Proc Natl Acad Sci U S A. 1975 Sep;72(9):3531-5. doi: 10.1073/pnas.72.9.3531.

Abstract

Genetic information from the bacterium Escherichia coli was transferred to human cells by means of the specialized transducing phage lambda plac carrying the bacterial z gene for the enzyme beta-galactosidase (geta-D-galactoside galactohydrolase, EC 3.2.1.23). As recipient cells, cultured skin fibroblasts from a patient with generalized gangliosidosis (GMI-gangliosidosis Type I) characterized by a severe deficiency of beta-galactosidase activity were used. The deficient human cells were incubated with the bacteriophage lambda plac or lambda plac DNA and beta-galactosidase activity was measured in order to detect gene transfer and acceptance of the prokaryotic information in the mammalian system for transcription and translation. The expression of the phage genome in the deficient fibroblasts could be demonstrated by detection of higher beta-galactosidase activity after incubation with phage lambda plac in three out of 19 experiments and in four out of 16 experiments after treatment with lambda plac DNA. Lambda plac DNA induced much higher enzyme activities than infective phage particles. Immunochemical and physicochemical assays could not distinguish the induced beta-galactosidase activity from that of the z-gene product of E. coli.

摘要

借助携带细菌β-半乳糖苷酶(β-D-半乳糖苷半乳糖水解酶,EC 3.2.1.23)的z基因的特异性转导噬菌体λplac,将来自大肠杆菌的遗传信息转移至人类细胞。作为受体细胞,使用了来自一名全身性神经节苷脂病(GM1-神经节苷脂病I型)患者的培养皮肤成纤维细胞,该患者表现为严重缺乏β-半乳糖苷酶活性。将缺陷的人类细胞与噬菌体λplac或λplac DNA一起孵育,并测量β-半乳糖苷酶活性,以检测基因转移以及原核信息在哺乳动物系统中进行转录和翻译的接受情况。在19次实验中的3次,用噬菌体λplac孵育后,以及在16次实验中的4次,用λplac DNA处理后,通过检测更高的β-半乳糖苷酶活性,可证明噬菌体基因组在缺陷成纤维细胞中的表达。λplac DNA诱导的酶活性比感染性噬菌体颗粒高得多。免疫化学和物理化学分析无法区分诱导的β-半乳糖苷酶活性与大肠杆菌z基因产物的活性。

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