Banville D, Williams J G
J Mol Biol. 1985 Aug 20;184(4):611-20. doi: 10.1016/0022-2836(85)90307-9.
We have analysed beta-globin mRNA sequences in total RNA extracted from embryos and tadpoles of Xenopus laevis at different stages of development and we have identified the most abundantly transcribed beta-globin mRNA (beta T1). The entire nucleotide sequence of a cDNA clone corresponding to this mRNA is known. We have now identified the gene corresponding to this mRNA and we have determined the nucleotide sequences of its immediate 5'-flanking region. Using a DNA fragment from within the coding region of the cloned beta T1 cDNA we show, by primer extension analysis, that beta T1 mRNA is first detectable at stage 28-32 of development. This is the time at which the first presumptive erythropoietic tissue, the ventral blood island, becomes observable histologically. We show that two minor beta-globin genes, distinct from beta T1, are expressed during early stages of development, and that their expression ceases shortly after the beginning of the feeding stage. We term these two early larval genes beta E1 and beta E2. A third minor beta-globin gene is expressed during early development but, unlike beta E1 and beta E2, it is also expressed throughout subsequent larval development. We term this gene beta T2 and show that it corresponds to a gene previously termed beta LII. Finally, using a primer derived from the major adult beta-globin gene (beta 1), we have analysed the accumulation of the major adult beta-globin mRNA during larval development, and we show that this sequence does not accumulate to any significant level before metamorphosis.
我们分析了从非洲爪蟾不同发育阶段的胚胎和蝌蚪中提取的总RNA中的β-珠蛋白mRNA序列,并鉴定出转录量最高的β-珠蛋白mRNA(βT1)。已知与该mRNA对应的cDNA克隆的完整核苷酸序列。我们现已鉴定出与该mRNA对应的基因,并确定了其紧邻的5'-侧翼区域的核苷酸序列。利用克隆的βT1 cDNA编码区内的一个DNA片段,通过引物延伸分析,我们发现βT1 mRNA在发育的第28 - 32阶段首次可检测到。这正是第一个假定的造血组织——腹侧血岛在组织学上变得可观察到的时间。我们发现,有两个与βT1不同的次要β-珠蛋白基因在发育早期表达,且它们的表达在摄食阶段开始后不久就停止了。我们将这两个早期幼虫基因命名为βE1和βE2。第三个次要β-珠蛋白基因在早期发育期间表达,但与βE1和βE2不同的是,它在随后的幼虫发育过程中也持续表达。我们将这个基因命名为βT2,并表明它对应于一个先前称为βLII的基因。最后,利用源自主要成年β-珠蛋白基因(β1)的引物,我们分析了主要成年β-珠蛋白mRNA在幼虫发育过程中的积累情况,结果表明该序列在变态前不会积累到任何显著水平。