Open AccessIsolation of Genomic DNA Controlling Mouse Melanoma Antigen Defined by Monoclonal Antibody
Author(s) -
Okitsu Akira,
Tagawa Masatoshi,
Tamura Yoshio,
Kanno Masamoto,
Matsubara Hisahiro,
Ito Toshihiro,
Imai Kenji,
Shigemoto Kazuhiro,
Nakamura Ikuro,
Koseki Haruhiko,
Lau YunFai,
Taniguchi Masaru
Publication year - 1988
Publication title -
japanese journal of cancer research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 0910-5050
DOI - 10.1111/j.1349-7006.1988.tb02228.x
Subject(s) - cosmid , biology , microbiology and biotechnology , monoclonal antibody , antigen , insert (composites) , cell sorting , genomic dna , transfection , dna , antibody , cell culture , genetics , flow cytometry , mechanical engineering , engineering
We have isolated the genomic DNA controlling the expression of murine specific melanoma antigen by employing cosmid shuttle vector and monoclonal antibody. Transfection of the cosmid library derived from mouse melanoma cells into human melanomas and repeated cell sortings of the fluorescence‐bright population enabled us to enrich the antigen‐positive transfectants. We rescued a 34.8 kb DNA fragment from the transfectants by in vitro packaging and showed it to be responsible for the antigen expression. However, we noticed instability of the antigen expression when the selection pressure imposed by the cell sorting was removed. This seemed to be due to the fact that the insert DNA was preferentially deleted from this cosmid vector without loss of the vector sequence itself.