Gaucher disease: retrovirus-mediated correction of the enzymatic defect in cultured cells.
Gaucher disease: retrovirus-mediated correction of the enzymatic defect in cultured cells.
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戈谢病：逆转录病毒介导的培养细胞酶促缺陷的校正。
DOI：
10.1101/sqb.1986.051.01.120
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Beutler,E
中科院分区：
文献类型：
--
作者：
Sorge,J;Kuhl,W;West,C;Beutler,E
方法DNA序列分析。如所述克隆人葡糖脑苷脂酶cDNA（索尔热等人，1985 a）。将DNA片段亚克隆到质粒pBR 322中，并通过Maxam和吉尔伯特0980的方法进行测序。这些研究中使用的小鼠单克隆抗体的生产和特性在其他地方有详细描述（Beutler et al. 1984）。将从美国典型培养物保藏中心获得的NIH-3 T3小鼠成纤维细胞在补充有5%小牛血清的Dulbecco改良Eagle培养基中培养。培养的皮肤成纤维细胞（指定为H）取自典型I型戈谢病患者。在逆转录病毒基因转移之前，用Epstein-Burr病毒使来自患有典型I型戈谢病的患者（指定为F）的淋巴细胞永生化。
METHODSDNA sequence analysis. The human glucocerebrosidase cDNA was cloned as described (Sorge et al. 1985a). DNA fragments were subcloned into the plusmid pBR322 and sequenced by the method of Maxam and Gilbert 0980).Cell culture. The production and characteristics of the mouse monoclonal antibodies used in these investigations are described in detail elsewhere (Beutler et al. 1984). NIH-3T3 mouse fibroblasts obtained from the American Type Culture Collection were cultured in Dulbecco's modified Eagle's medium supplemented with 5% calf serum. Cultured skin fibroblasts (designated H) were obtained from a patient with typical type I Gaucher disease. Lymphocytes from a patient (designated F) with typical type I Gaucher disease were immortalized with Epstein-Burr virus prior to retrovirus gene transfer.