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Measurement of histone mRNA transcript abundance in Xenopus oocytes by a quantitative primer extension assay
Author(s) -
BrashearsMacatee Sarah,
Hinkley Craig,
Perry Michael
Publication year - 1990
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080250105
Subject(s) - biology , xenopus , primer extension , primer (cosmetics) , microbiology and biotechnology , histone , gene , messenger rna , oogenesis , gene expression , oocyte , transcription (linguistics) , genetics , embryo , linguistics , chemistry , philosophy , organic chemistry
A quantitative primer extension method was used to measure the mass of histone gene transcripts in mature oocytes of the amphibian Xenopus laevis. The procedure, using a large excess of gene‐specific oligonucleotide primer and continuous incorporation of a radiolabeled deoxynucleoside triphosphate precursor, is more sensitive and quantitative than primer extension assays employing end‐labeled primers. It was determined that there are stoichometric amounts, approximately 2 × 10 8 copies, of mRNA for each of the five major histone gene classes in mature Xenopus oocytes. These observations are consistent with a model whereby transcription of these genes is coordinately regulated in a cell cycle‐independent manner during amphibian oogenesis.