Kubo H, Kawano T, Tsubuki S, Kotani M, Kawasaki H, Kawashima S
Department of Membrane Biochemistry, Tokyo Metropolitan Institute of Medical Science, Japan.
Dev Growth Differ. 2000 Aug;42(4):419-27. doi: 10.1046/j.1440-169x.2000.00526.x.
The egg envelope is a kind of extracellular matrix, which surrounds growing oocytes, ovulated eggs and early embryos. Among the glycoprotein components of the Xenopus laevis egg envelope, gp43/gp41 and gp69/64 have already been shown to be frog homologs of the mammalian zona pellucida components ZP3 and ZP2, respectively. To determine the structure of another major component of egg envelope, gp37, the peptides isolated from the lysyl endopeptidase digests of gp37 were sequenced for amino acids to design degenerate primers for polymerase chain reaction. By reverse transcription-polymerase chain reaction with a poly(A)+ RNA from the ovary of a postovulated female Xenopus, a specifically amplified band was obtained and sequenced. The upstream and downstream sequences of the sequenced region were completed by 5'- and 3'-rapid amplification of cDNA ends, respectively. The gp37 cDNA comprises 1674 bp and contains one open reading frame encoding a polypeptide with 543 amino acids. The predicted amino acid sequence of the gp37 cDNA has a close similarity to that of mammalian ZP1. Northern blot and in situ hybridization studies indicated that the transcript (1.8 kb) is exclusively expressed in the oocytes, particularly in the previtellogenic young oocytes, just like the expression pattern of gp43 mRNA, suggesting a coordinate transcription of the gp43 and gp37 genes in Xenopus.
卵膜是一种细胞外基质,它围绕着生长中的卵母细胞、排卵后的卵子和早期胚胎。在非洲爪蟾卵膜的糖蛋白成分中,gp43/gp41和gp69/64已分别被证明是哺乳动物透明带成分ZP3和ZP2的蛙类同源物。为了确定卵膜的另一种主要成分gp37的结构,对从gp37的赖氨酰内肽酶消化产物中分离出的肽段进行氨基酸测序,以设计用于聚合酶链反应的简并引物。通过对排卵后雌性非洲爪蟾卵巢的聚腺苷酸加尾RNA进行逆转录聚合酶链反应,获得了一条特异性扩增带并进行了测序。测序区域的上游和下游序列分别通过5'-和3'-cDNA末端快速扩增完成。gp37 cDNA包含1674 bp,含有一个开放阅读框,编码一个含有543个氨基酸的多肽。gp37 cDNA的预测氨基酸序列与哺乳动物ZP1的氨基酸序列非常相似。Northern印迹和原位杂交研究表明,该转录本(1.8 kb)仅在卵母细胞中表达,特别是在卵黄生成前的幼卵母细胞中,就像gp43 mRNA的表达模式一样,这表明非洲爪蟾中gp43和gp37基因存在协同转录。