鸡胚发育过程中组蛋白H5和珠蛋白mRNA的分析。

Analysis of histone H5 and globin mRNAs in chick-embryo development.

作者信息

Colman A, Krieg P, Wells J R

出版信息

Eur J Biochem. 1983 Apr 5;131(3):551-4. doi: 10.1111/j.1432-1033.1983.tb07297.x.

DOI:10.1111/j.1432-1033.1983.tb07297.x

Abstract

Transcripts from histone H5 and alpha and beta chicken globin genes were analysed in total RNA extracted from 2-6-day-old chick embryos. Denatured RNA, separated on gels, was transferred to either aminophenylthioether (APT) paper or to nitrocellulose and was detected with 32P-labelled recombinant DNA probes. Good resolution was achieved with methyl mercury hydroxide gels and aminophenylthioether transfer, allowing estimation of H5 and alpha and beta globin mRNA sizes. With glyoxal gels and nitrocellulose transfer, alpha and beta globin mRNAs were not resolved, but an increased sensitivity of detection of at least 10-fold was achieved. Levels of H5 and alpha globin mRNAs increased in parallel over 2-6 days. The appearance of definitive red cells in 6-day embryos was indicated by detection of adult beta globin transcripts at this stage.

摘要

对从2至6日龄鸡胚中提取的总RNA进行了组蛋白H5以及α和β鸡珠蛋白基因转录本的分析。变性RNA在凝胶上分离后，转移至氨苯基硫醚（APT）纸或硝酸纤维素膜上，并用32P标记的重组DNA探针进行检测。用氢氧化甲基汞凝胶和氨苯基硫醚转移法可实现良好的分辨率，从而能够估算H5以及α和β珠蛋白mRNA的大小。使用乙二醛凝胶和硝酸纤维素膜转移法时，α和β珠蛋白mRNA无法分辨，但检测灵敏度提高了至少10倍。在2至6天内，H5和α珠蛋白mRNA的水平平行升高。通过在这个阶段检测到成年β珠蛋白转录本，表明6日龄胚胎中出现了成熟红细胞。