Measurement of histone mRNA transcript abundance in Xenopus oocytes by a quantitative primer extension assay.
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 stoichiometric amounts, approximately 2 X 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.
Brashears-Macatee
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S
Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Hinkley
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C
Perry
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M
D000818:Animals; D005260:Female; D005784:Gene Amplification; D006657:Histones / Q000235:genetics*; D009841:Oligonucleotides; D009865:Oocytes / Q000378:metabolism; D009866:Oogenesis / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D014158:Transcription, Genetic; D014982:Xenopus laevis / Q000235:genetics*
D006657:Histones; D009841:Oligonucleotides; D012333:RNA, Messenger