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Measurement of histone mRNA transcript abundance in Xenopus oocytes by a quantitative primer extension assay.

作者信息

Brashears-Macatee S, Hinkley C, Perry M

机构信息

Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.

出版信息

Mol Reprod Dev. 1990 Jan;25(1):22-7. doi: 10.1002/mrd.1080250105.

Abstract

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.

摘要

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