| Bone morphogenetic proteins (BMPs) are important members of the transforminggrowth factor β (TGF-β) super family. In recent years, with the development of the researchresults, we have found the function of BMPs not only in the bone tissue directionaldifferentiation, but also in the mesodem induction, physical development, tooth development,formation and animal reproduction character expression. On the basis of previous work, theBMPs cloning sequencing and analysis of New Zealand white rabbits were studied indifferent development periods and spatio-temporal difference expression in different tissue, inorder to determine the role of different development stages of BMPs in rabbits from theperspective of gene expression spectrum regularity. According to GenBank of released pig,cattle, sheep and mice bone morphogenetic protein2(BMP-2) and bone morphogeneticprotein (BMP-4) sequence in NCBI, and use computer software DNAMAN to find out BMP-2gene and BMP-4gene cDNA of conservative area, design specific primers, using RT-PCRtechnology for gene cloning, then cloning sequence as a probe to continue electronic cloning,using bioinformatics software on the molecular structure and characteristics of two kinds ofgenetic analysis and prediction, and then apply real-time quantitative polymerase chainreaction (PCR) to detect the mRNA level of two genes in different development period of theNew Zealand white rabbits’ ovaries, uterus, fallopian tubes, kidneys, liver, spleen andduodenum tissue expression. The major results are as follows:1. Part of cDNA fragment of transforming growth factor super family member of rabbitBMP-2and BMP-4gene by RT-PCR from ovaries, uterus, fallopian tubes were cloned. Theresult showed the length of BMP-2is299bp, the length of BMP-4is175bp.2. Using partial cDNA of sequenced BMP-2and BMP-4as the probe, then search for thenr database and EST database, BMP-2of1150bp and BMP-4of920bp were respectivelycloned.3. The rabbit electronic cloning of BMP-2and BMP-4by the bioinformatics wereanalyzed, the full-length BMP-2CDS is1149bp, encoding383amino acids. The length ofsequence of BMP-4is918bp and encoding306amino acids.4. The homology of BMP-2and BMP-4gene were analyzed by the bioinformaticsmethods and related software: the nucleotide identity of New Zealand rabbit BMP-2genewith sheep, cattle, pig, mouse, rat, human is86,86,86,93,93,87percent respectively. Thenucleotide identity of BMP-4gene with sheep, cattle, pig, mouse, rat is93,94,94,91,92 percent respectively.5. By real-time quantitative PCR, New Zealand rabbit BMP-2and BMP-4were detected,the relative expression of BMP-2gene at the30day age,60day age and180day age ishigher in ovarian tissue, the liver tissue of120day age is highest, the relative expression ofBMP-4gene of ovarian at60day and180day is higher, duodenum of30day is higher, liverof pregnancy rabbit is higher.6. Two genes in the same tissue of different growth period, the expression of regularityis shown as: the mRNA of ovariesof BMP-2ranged largely,60days of age reached the peak,the expression of oviduct, spleen and uterus are the highest in120days of age. Ovaries inBMP-4mRNA expression of the highest levels within60days of age, the oviduct, uterus,kidney, liver, spleen, express the highest levels in the gestational6day rabbit, duodenalexpress the highest levels within30days of age.7. BMP-2gene and BMP-4gene in the ovarian tissue has a consistent expression trend,namely the highest60days of age, after the fall, to180days of age rise again. Expression inthe duodenum also exist obvious similarity, namely with the increase of age, the expressionof both genes present a downward trend. |
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