Wegener G, Kläner S, Sauer H W
Physiologischer Lehrstuhl, Zoologisches Institut der Universität Heidelberg, Deutschland.
Wilhelm Roux Arch Entwickl Mech Org. 1971 Jun;167(2):118-136. doi: 10.1007/BF00577036.
Proteins in homogenates of yolk-rich cricket eggs were fractionated by sonication, differential centrifugation, acetic acid-extraction and column chromatography on hydroxyapatite and separated by SDS-polyacrylamide electrophoresis in 8 M urea. Radioactivity was introduced by incubating the eggs, which are impermeable to labeled amino acids, in aCO-atmosphere.C-label in protein extracts was resistant to hot TCA-extraction and was proved to be mainly in glutamic acid and aspartic acid. The profiles of radioactivity across the electropherogram as analysed by gel-fractionation and scintillation counting or by radioautography were found to be distinct from the pattern of stained protein-bands. During early development characteristic changes in the patterns of radioactive proteins occurred at the three stages tested: cleavage (omnipotency), germ-disc (determination) and germ-band (primary differentiation).
富含卵黄的蟋蟀卵匀浆中的蛋白质通过超声处理、差速离心、乙酸提取和羟基磷灰石柱色谱进行分级分离,并在8M尿素中通过SDS-聚丙烯酰胺凝胶电泳进行分离。通过将对标记氨基酸不可渗透的卵在含14CO2的气氛中孵育来引入放射性。蛋白质提取物中的14C标记对热三氯乙酸提取具有抗性,并且被证明主要存在于谷氨酸和天冬氨酸中。通过凝胶分级分离和闪烁计数或放射自显影分析的电泳图谱上的放射性分布被发现与染色蛋白带的模式不同。在早期发育过程中,在所测试的三个阶段(卵裂期(全能性)、胚盘期(决定)和胚带期(初步分化)),放射性蛋白质的模式发生了特征性变化。