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Application and Limitations of Differential Hybridization in the Isolation of T Cell‐Specific cDNA Clones
Author(s) -
Shigemoto Kazuhiro,
Kanno Masamoto,
Sakamoto Tohru,
Imai Kenji,
Taniguchi Masaru
Publication year - 1987
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1987.tb03151.x
Subject(s) - complementary dna , biology , microbiology and biotechnology , northern blot , southern blot , cloning (programming) , dot blot , nucleic acid thermodynamics , differential display , hybridization probe , cdna library , molecular cloning , gene , genetics , rna , computer science , programming language
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 × 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.