Kovesdi I, Smith M J
Mol Cell Biol. 1985 Nov;5(11):3001-8. doi: 10.1128/mcb.5.11.3001-3008.1985.
Actin coding sequence cDNA probes were used to quantitate the number of transcripts in RNA from eggs, embryos, and tube feet of the sea star Pisaster ochraceus. Transcript concentrations were measured in both total RNA and in poly(A)+ RNA by titration and hybridization kinetic methods. Surprisingly, the actin transcript number in sea star eggs is two orders of magnitude greater than in sea urchin eggs. There are at least 2.9 X 10(5) actin transcripts per sea star egg, 1.2 X 10(5) per 48-h gastrula and 1.9 X 10(5) per 72-h gastrula. The number of actin transcripts per unit mass of extracted tube foot RNA is lower than in developmental stages. The relative abundance and size of actin transcripts was determined by Northern and dot blot analyses using probes containing actin coding DNA or 3'-untranslated-region sequences. The actin transcript in eggs and embryos is 2,300 nucleotides (nt) long and originates from the Cy (cytoplasmic) gene class. In tube feet, the most abundant actin transcript is 2,200 nt long and originates from the M (muscle) gene class. Tube feet also contain, at lower abundance, 2,300-nt transcripts of the Cy gene type expressed in eggs and embryos.
肌动蛋白编码序列cDNA探针被用于定量分析海星海盘车卵子、胚胎和管足RNA中的转录本数量。通过滴定和杂交动力学方法测量了总RNA和聚腺苷酸加尾RNA(poly(A)+ RNA)中的转录本浓度。令人惊讶的是,海星卵子中的肌动蛋白转录本数量比海胆卵子中的高两个数量级。每个海星卵子至少有2.9×10⁵个肌动蛋白转录本,每48小时的原肠胚中有1.2×10⁵个,每72小时的原肠胚中有1.9×10⁵个。每单位质量提取的管足RNA中的肌动蛋白转录本数量低于发育阶段。使用包含肌动蛋白编码DNA或3'非翻译区序列的探针,通过Northern印迹和斑点印迹分析确定了肌动蛋白转录本的相对丰度和大小。卵子和胚胎中的肌动蛋白转录本长度为2300个核苷酸(nt),源自Cy(细胞质)基因类别。在管足中,最丰富的肌动蛋白转录本长度为2200 nt,源自M(肌肉)基因类别。管足中还含有较低丰度的、在卵子和胚胎中表达的2300 nt的Cy基因类型转录本。