Deyev S M, Urakov D N, Stepchenko A G, Polanovsky O L
V.A. Engelhardt Institute of Molecular Biology, Academy of Sciences of USSR, Moscow.
Genetica. 1991;85(1):45-51. doi: 10.1007/BF00056105.
The hybridoma PTF-02 secretes an antibody against pig transferrin. Rearranged genes for heavy and light immunoglobulin chains have been studied in the genomes of this hybridoma and in the parent myeloma P3-X63.Ag8.653. The hybridoma was shown to contain three rearranged allelic variants of the heavy chain gene's locus. The gene H2, responsible for synthesis of the heavy chain of the antibody to transferrin, was transmitted in the hybridoma cell from a lymphocyte. Two other genes (H1 and H3) were found both in the hybridoma and parent myeloma genomes. The gene H1 was identified in MOPC21 myeloma, which is a precursor of the X63.Ag8 descendent line. Rearranged k genes were also identified both in the hybridoma and parent myeloma. A functional (K2) gene and a fetal (F) gene appeared in the hybridoma genome from an antigen-stimulated normal lymphocyte. The fetal gene was lost in the course of continuous cultivation of the hybridoma PTF-02 cell line. The gene K1 was transmitted from the myeloma used for fusion. In such a way, the pedigree of rearranged heavy and light chain genes in the hybridoma PTF-02 was established. The results obtained in this work may be relevant to many hybridomas whose immortalizing fusion partner is a MOPC21 derivative, and allow one to identify and isolate functional variable genes to create recombinant constructions.
杂交瘤PTF - 02分泌一种抗猪转铁蛋白的抗体。已在该杂交瘤的基因组以及亲本骨髓瘤P3 - X63.Ag8.653的基因组中研究了重链和轻链免疫球蛋白基因的重排。结果表明,该杂交瘤含有重链基因座的三种重排等位基因变体。负责合成转铁蛋白抗体重链的H2基因是从淋巴细胞传递到杂交瘤细胞中的。另外两个基因(H1和H3)在杂交瘤和亲本骨髓瘤基因组中均有发现。H1基因在MOPC21骨髓瘤中被鉴定出来,MOPC21骨髓瘤是X63.Ag8后代细胞系的前体。在杂交瘤和亲本骨髓瘤中也鉴定出了重排的κ基因。一个功能性(K2)基因和一个胎儿型(F)基因来自抗原刺激的正常淋巴细胞,出现在杂交瘤基因组中。在杂交瘤PTF - 02细胞系的连续培养过程中,胎儿型基因丢失了。K1基因是从用于融合的骨髓瘤中传递而来的。通过这种方式,确定了杂交瘤PTF - 02中重链和轻链基因重排的谱系。这项工作所获得的结果可能与许多永生融合伙伴是MOPC21衍生物的杂交瘤相关,并有助于鉴定和分离功能性可变基因以构建重组体。