关键词: 3′RACE, 3′非翻译序列, 克隆, BMP7, 家兔

Abstract:

On the basis of known partial coding DNA sequence (GenBank accession No. AF413111), we tried to clone the full mRNA of rabbit BMP7 gene by sub-cloning and RACE methods. Then after, bioinformatic analysis on the acquired sequences were conducted. The sequencing results showed that the all cloned sequences could be assembled into a 1 654 bp long DNA fragment, which contained the near full coding cDNA for the propetide, full coding cDNA for the mature protein, and full length of 3′UTR of rabbit BMP7. The newly cloned sequences extended the 5′and 3′ends of known partial DNA sequence by 395 bp and 628 bp, respectively. Sequence comparisons revealed that the full length of coding cDNA of rabbit BMP7 was 91.89% and 89.32% identical to that of human and mouse, while the deduced amino acids was 96.51% and 96.01% identical, respectively. The 3′UTR of rabbit BMP7 was 446 bp in length, 57.38% and 45.57% identical to that of human or mouse, respectively. In addition, there were two closely arranged AATAAA sites in the 3′UTR of rabbit BMP7 gene, as well as in human BMP7. The deduced mature protein of rabbit BMP7 had the seven conservative cysteines and a TGF-b family signature, which were the characters of all BMPs. Our results suggested that the main part of rabbit BMP7 had been successively cloned. The alternation of polyadenization sites in the 3′UTR of rabbit BMP7 maybe related to the posttranscriptional regulation of the gene.