IMMUNOGLOBULIN GENE-EXPRESSION IN TRANSFORMED LYMPHOID-CELLS
IMMUNOGLOBULIN GENE-EXPRESSION IN TRANSFORMED LYMPHOID-CELLS
复制标题
DOI：
10.1073/pnas.80.3.825
复制
发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
BERG, P
中科院分区：
文献类型：
--
作者：
OI, VT;MORRISON, SL;BERG, P
用质粒载体pSV 2-gpt成功地转染了大肠杆菌黄嘌呤-鸟嘌呤磷酸核糖转移酶基因（gpt）到骨髓瘤、杂交瘤和胸腺瘤细胞系中。转化的细胞合成细菌酶5-磷酸-α-D-核糖-1-二磷酸：黄嘌呤磷酸核糖基转移酶（XGPRT; EC 2.4.2.22），并在选择性培养基中保持4个月以上。通过用含有来自S107骨髓瘤细胞系的重排的K轻链基因组片段的pSV 2-gpt转染细胞，获得表达K免疫球蛋白轻链的类肉瘤细胞系。S107轻链在gpt转化的J558 L骨髓瘤细胞中合成，并且通过免疫沉淀和二维凝胶电泳判断，与S107骨髓瘤细胞系合成的轻链相同。此外，该轻链作为完整抗体分子的一部分由转化的杂交瘤细胞合成和分泌，所述转化的杂交瘤细胞通常分泌IgGl（γ，κ）抗体分子。在类似转化的大鼠骨髓瘤或小鼠胸腺瘤系中未检测到轻链合成。
Myeloma, hybridoma, and thymoma cell lines have been successfully transfected for the Escherichia coli xanthine-guanine phosphoribosyltransferase gene (gpt) by using the plasmid vector pSV2-gpt. The transformed cells synthesize the bacterial enzyme 5-phospho-alpha-D-ribose-1-diphosphate:xanthine phosphoribosyltransferase (XGPRT; EC 2.4.2.22) and have been maintained in selective medium for over 4 months. Lymphoid cell lines expressing a K immunoglobulin light chain were obtained by transfecting cells with pSV2-gpt containing a rearranged K light chain genomic segment from the S107 myeloma cell line. The S107 light chain is synthesized in gpt-transformed J558L myeloma cells and is identical to the light chain synthesized by the S107 myeloma cell line, as judged by immunoprecipitation and two-dimensional gel electrophoresis. Furthermore, this light chain is synthesized and secreted as part of an intact antibody molecule by transformed hybridoma cells that normally secrete an IgGl (gamma, K) antibody molecule. No light chain synthesis was detected in a similarly transformed rat myeloma or a mouse thymoma line.