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差异杂交在T细胞特异性cDNA克隆分离中的应用及局限性

Application and limitations of differential hybridization in the isolation of T cell-specific cDNA clones.

作者信息

Shigemoto K, Kanno M, Sakamoto T, Imai K, Taniguchi M

机构信息

Department of Immunology, School of Medicine, Chiba University.

出版信息

Microbiol Immunol. 1987;31(9):899-909. doi: 10.1111/j.1348-0421.1987.tb03151.x.

Abstract

We investigated the limitations and effectiveness of differential hybridization in the cloning of T cell-specific cDNA (complementary DNA) molecular clones. By using the technique with T cell and B cell cDNA probes, together with Northern blot analysis, we successfully isolated cDNA clones exclusively expressed in T cells from 1 X 10(4) plaque-forming units of a T cell hybridoma. These clones represent 0.068% of the mass of the cytoplasmic mRNA. Our result shows that differential hybridization is an effective procedure when used in combination with Northern blot analysis for screening of genes selectively expressed in T cells.

摘要

我们研究了差异杂交在克隆T细胞特异性互补DNA(cDNA)分子克隆中的局限性和有效性。通过将该技术与T细胞和B细胞cDNA探针一起使用,并结合Northern印迹分析,我们成功地从1×10⁴个T细胞杂交瘤的噬斑形成单位中分离出仅在T细胞中表达的cDNA克隆。这些克隆占细胞质mRNA总量的0.068%。我们的结果表明,差异杂交与Northern印迹分析结合使用时,是筛选在T细胞中选择性表达基因的有效方法。

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