Lasky L A, Lev Z, Xin J H, Britten R J, Davidson E H
Proc Natl Acad Sci U S A. 1980 Sep;77(9):5317-21. doi: 10.1073/pnas.77.9.5317.
mRNA prevalence during sea urchin development was measured by treating cDNA clone colonies with labeled cDNAs transcribed from unfertilized egg and embryo poly(A)-RNAs. The number of cytoplasmic transcripts per embryo complementary to several clones was determined independently by titration with poly(A)-RNA in solution, and the amount of cDNA bound to these clones in colony hybridizations was shown to be proportional to the concentration of the respective poly(A)-RNAs in the embryo cytoplasm. At the gastrula stage, the most prevalent mRNA species occur in about 10(6) molecules per embryo. If all cells were equivalent, this would be a few hundred molecules per cell. By pluteus stage, the prevalence of some sequences has increased more than 10-fold. Most, though not all, sequences prevalent in later embryos are also present in the maternal RNA of the unfertilized egg. For most poly(A)-RNA sequences, the prevalence levels determined during oogenesis are maintained through the pluteus stage, whereas a minority of sequences display sharp stage-specific changes in representation during development.
通过用从未受精卵和胚胎多聚腺苷酸(poly(A))RNA转录的标记cDNA处理cDNA克隆菌落,来测量海胆发育过程中mRNA的丰度。通过用溶液中的多聚腺苷酸(poly(A))RNA滴定,独立测定每个胚胎中与几个克隆互补的细胞质转录本数量,并且在菌落杂交中与这些克隆结合的cDNA量显示与胚胎细胞质中各自多聚腺苷酸(poly(A))RNA的浓度成比例。在原肠胚阶段,最丰富的mRNA种类每个胚胎中约有10⁶个分子。如果所有细胞都是等同的,那么每个细胞将有几百个分子。到长腕幼虫阶段,一些序列的丰度增加了10倍以上。大多数(尽管不是全部)在后期胚胎中丰富的序列也存在于未受精卵的母体RNA中。对于大多数多聚腺苷酸(poly(A))RNA序列,在卵子发生过程中确定的丰度水平在长腕幼虫阶段之前保持不变,而少数序列在发育过程中表现出明显的阶段特异性表达变化。