| Bone morphogenetic proteins (BMPs) and growth differentiation factors (GDFs) are important members of the transforming growth factorβ(TGF-β) super family. They are essential for animal reproductive performance, especially for regulating the growth of follicles and animal reproductive function. So, the study on molecule clone and expression of the BMPs, GDFs and their receptors in different organizations is to make sure the reproductive function of BMPs and their receptors in different reproductive stages, and to clarify the sterility or fetal mechanism from gene angle. In the present study, based on the findings of other species (human, cow, pig, mouse, chicken, et al) bone morphogenetic proteins, we cloned the four genes of related reproductive function gene of Mongolian cattle by RT-PCR. The characteristics and the molecular structure of the four genes were analyzed and predicted by the Bioinformatics. Then we examined the mRNA level expression of the four genes in bovine ovary, uterus, oviduct, and kidney by semi-quantitative RT-PCR and In situ hybridization. The major results are as follows:1. Part of cDNA fragment of transforming growth factor super family members of bovine BMP15, BMP-IB, GDF9, and BMP4 gene by RT-PCR from ovaries were cloned and the result showed a high degree of sequence homology to each other mammalian sequence at both the nucleotide and peptide levels, conforming to the characteristics of TGF-βfamily members.2. The 3′sequences of the Mongolian cattle BMP15 CDS, BMPR-IB and BMP4 by the Bioinformatics were analyzed, the full-length BMP15 CDS is 1185 bp, including two extrons and encoding 394 amino acids. The sequence has been registered in the GenBank, and the number is DQ463368; The length of 3′sequences of BMPR-IB gene is 953 bp and encoding 302 amino acids. The sequence has been registered in the GenBank, and the number is DQ489533; The full-length of 3′sequences of BMP4 gene was cloned by RACE technology, the length is 1306 bp and encoding 389 amino acids, including the 121 bp length of the 3′non-translated head and the PolyA that contain 15 bases, the sequence has been registered in the GenBank, and the number is AY854957; Cloned bovine GDF9 gene conservative region and the length is 264 bp.3. The homology of BMP15, BMPR-IB, and BMP4 gene were analyzed by the bioinformatics methods and related softwares: the nucleotide identity of bovine BMP15 gene with sheep, pig, zebrafish is 98.5, 87.6, 29.1 percent respectively. In the 291 ~ 394 amino acids, exist a conservative region is the homologous or heterologous domain of all the TGF-βfamily members by forecast bovine BMP15 protein conservative region and bovine BMP15 amino acid sequence exist four functional sites.Bovine BMPR-IB gene encode 302 amino acid sequences and the nucleotide sequence with the human and poultry are highly conservative. The nucleotide identity of bovine BMP-IB cDNA genes with typical eukaryote of sheep, goat, human, monkey, pig, chicken is more than 90%.The cDNA sequence contains 1170 bp coding region and the 3'terminal amino acid sequences includes seven cysteines which is the character of BMP gene in the analysis of bovine BMP4 gene sequence. The nucleotide identity of bovine BMP4 gene with human, sheep, mice, rats, dogs, chicken is 90.2, 94.2, 90.9, 90.4, 85.4, 73.6 percent respectively.4. The four genes mRNA expression in bovine ovary, uterus, oviduct, and kidney were examined by semi-quantitative RT-PCR. The four genes all have been expressed in the bovine ovary, uterus, oviduct, and kidney, but different genes in different tissues have different expression levels.5. The BMP15 gene mRNA expression in ovary were examined by In situ hybridization and the results showed that the BMP15 mRNA expression in bovine ovaries, and the location differed due to different follicle developing stage: in the primary and early secondary follicles, signals were located mainly in granulose cells, and in the late secondary follicles, signals were located solely in zona pellucida.
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