Bhatia R, Dube D K, Lemanski L F
SUNY Health Science Center at Syracuse, Department of Anatomy and Cell Biology 13210, USA.
Biochem Mol Biol Int. 1996 May;38(6):1079-85.
We have isolated and sequenced a full-length (0.9 kb) cDNA clone of ribosomal protein S3 by subtraction hybridization using a single-stranded cDNA library from stage 25-27 (tracer) and the mRNA from stage 15-17 (driver) of embryonic Mexican axolotl (Ambystoma mexicanum). The axolotl is a unique animal model for studying heart development as well as myofibrillogenesis because it carries a mutation in gene c. The deduced amino acid sequence of axolotl S3 protein shows about 93.9% identity with human S3 protein over a 243 amino acid residue overlap. When compared with mouse and Xenopus laevis ribosomal S3 proteins, the axolotl sequenc shows 94.3 and 93.9% identity respectively. Interestingly, the axolotl S3 sequence shows higher identity at the nucleic acid level with human and/or other mammals than with Xenopus. The S3 transcript, as determined by RT-PCR, is present at stage 2-4 in a lower amount and the onset of transcription is most likely at the beginning of gastrulation (10-12). The expression level of S3 transcripts reaches a maximum by mid gastrulation (stages 13-14) and then follows a biphasic pattern being lower at stages 16-17 with subsequently steady increases until the mid tailbud stages (25-27).
我们通过扣除杂交法,利用墨西哥钝口螈(Ambystoma mexicanum)胚胎第25 - 27阶段的单链cDNA文库(示踪剂)和第15 - 17阶段的mRNA(驱动子),分离并测序了核糖体蛋白S3的全长(0.9 kb)cDNA克隆。钝口螈是研究心脏发育以及肌原纤维形成的独特动物模型,因为它携带基因c的突变。钝口螈S3蛋白推导的氨基酸序列在243个氨基酸残基重叠区域与人类S3蛋白显示出约93.9%的同一性。与小鼠和非洲爪蟾核糖体S3蛋白相比,钝口螈序列分别显示出94.3%和93.9%的同一性。有趣的是,钝口螈S3序列在核酸水平上与人类和/或其他哺乳动物的同一性高于与非洲爪蟾的同一性。通过RT-PCR确定,S3转录本在第2 - 4阶段含量较低,转录起始最有可能在原肠胚形成开始时(第10 - 12阶段)。S3转录本的表达水平在原肠胚中期(第13 - 14阶段)达到最大值,然后呈现双相模式,在第16 - 17阶段较低,随后稳步增加,直到尾芽中期(第25 - 27阶段)。