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Isolation of α‐fetoprotein Messenger RNA
Author(s) -
MIURA K.,
LAW S. W.,
TAMAOKI T.
Publication year - 1978
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1978.tb03924.x
Subject(s) - messenger rna , polysome , microbiology and biotechnology , yolk sac , immunoprecipitation , agarose , rna , sepharose , affinity chromatography , chemistry , translation (biology) , biology , biochemistry , embryo , ribosome , gene , enzyme
Messenger RNA for α‐fetoprotein (AFP mRNA) was isolated from mouse yolk sac by a combination of indirect immunoprecipitation of AFP‐synthesizing polysomes and poly(U)‐Sepharose affinity chromatography of poly(A)‐containing mRNA. Over 200‐fold purification was achieved. Purified AFP mRNA migrated as a single band at about 21S on agarose gels in the presence of 6 M urea. Upon translation in wheat germ extracts, 98% of the protein synthesized reacted with specific antibodies to AFP. The total translation product was characterized by means of gel electrophoresis under two different conditions and by tryptic peptide analysis. It was concluded that the AFP preparation was essentially free of other yolk sac mRNA and was able to direct the synthesis of authentic AFP.