Purification of pre-prolactin mRNA from bovine anterior pituitary glands.
Total cytoplasmic RNA, isolated from bovine anterior pituitary glands, directed the synthesis of primarily two proteins in a wheat germ cell-free system. Analysis of wheat germ cell-free products by sodium dodecyl sulfate-slab gel electrophoresis, direct immunoprecipitation with prolactin aatibody, and prolactin- and growth hormone-specific radioimmunoassay indicated that the two proteins were pre-prolactin and pre-growth hormone. Agarose-urea-gel electrophoresis of poly(A)-containing RNA revealed the presence of a large symmetrical RNA peak with an estimated size of approximately 15 S. The RNA contained in this peak was eluted from the gel and translated in the wheat germ cell-free system. Analysis of the translation products by electrophoresis, direct immunoprecipitation, and radioimmunoassay resulted in the detection of only pre-prolactin. The extent of pre-prolactin mRNA purification was assessed by synthesizing DNA complementary to gel-purified pre-prolactin mRNA and then following the hybridization of this cDNA to gel purified pre-prolactin mRNA. The result of this hybridization is consistent with the cell-free translation studies, namely that pre-prolactin mRNA is the predominant mRNA component of pituitary poly(A)-containing RNA.
D000818:Animals; D002417:Cattle; D008970:Molecular Weight; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D010944:Plants / Q000378:metabolism; D011061:Poly A / Q000032:analysis; D011388:Prolactin / Q000096:biosynthesis*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D014908:Triticum / Q000378:metabolism
D011498:Protein Precursors; D012333:RNA, Messenger; D011061:Poly A; D011388:Prolactin