非洲爪蟾骨形态发生蛋白-1编码cDNA在胚胎发育早期的克隆与表达

Cloning and expression of cDNA encoding Xenopus laevis bone morphogenetic protein-1 during early embryonic development.

作者信息

Maéno M, Xue Y, Wood T I, Ong R C, Kung H F

机构信息

Laboratory of Biochemical Physiology, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702-1201.

出版信息

Gene. 1993 Dec 8;134(2):257-61. doi: 10.1016/0378-1119(93)90103-a.

DOI:10.1016/0378-1119(93)90103-a

PMID:8262384

Abstract

The Xenopus laevis DNA fragment encoding a protein homologous with human bone morphogenetic protein-1 (BMP-1) was amplified by polymerase chain reaction (PCR) from cDNA derived from stage 26 (st.26) embryonic RNA. Subsequently this fragment was used as a probe to isolate cDNA clones by screening of a X. laevis st.24 embryonic cDNA library. Two partial clones (22 and 63) were obtained and the missing 5'-end of the clone 22 was extended by the anchored PCR technique. The nucleotide sequence of the resulting clone (22AN) contained an open reading frame coding for a protein with 707 deduced amino acids. Three sizes of mRNA (2.9, 5.2 and 6.6 kb) were detected in blastula (st.9) and early gastrula (st.10) embryos, and in hatched tadpole (st.40), but little or no expression was observed in morula (st.7) and late gastrula (st.12) embryos, suggesting a physiological role(s) of X.laevis BMP-1 in normal embryonic development.

摘要

通过聚合酶链反应（PCR），从第26期（st.26）胚胎RNA衍生的cDNA中扩增出非洲爪蟾编码与人骨形态发生蛋白-1（BMP-1）同源蛋白质的DNA片段。随后，该片段用作探针，通过筛选非洲爪蟾st.24胚胎cDNA文库来分离cDNA克隆。获得了两个部分克隆（22和63），并通过锚定PCR技术延伸了克隆22缺失的5'端。所得克隆（22AN）的核苷酸序列包含一个开放阅读框，编码一个推导有707个氨基酸的蛋白质。在囊胚期（st.9）、早期原肠胚期（st.10）胚胎以及孵化后的蝌蚪（st.40）中检测到三种大小的mRNA（2.9、5.2和6.6 kb），但在桑椹胚期（st.7）和晚期原肠胚期（st.12）胚胎中几乎未观察到表达，这表明非洲爪蟾BMP-1在正常胚胎发育中具有生理作用。