摘要
在对已知部分编码序列(CDS)进行分析的基础上,采用RT-PCR分步扩增以及RACE方法,对家兔BMP7基因3′和5′末端未知序列进行了克隆与生物信息学分析。测序结果综合分析表明,所获序列共计1654bp,包括家兔BMP7近全长前肽、全长成熟肽CDS及3′非翻译序列(3′UTR),将已有的序列向5′和3′端分别延伸了395bp和628bp。序列对比表明,克隆的家兔BMP7 CDS部分与人、小鼠的对应序列的同源性分别为91.89%和89.32%,预测的氨基酸序列同源性分别为96.51%和96.01%。家兔BMP7 3′UTR长446bp,与人、小鼠对应序列同源性分别为57.38%和45.57%;具有2个转录终止信号位点。推测家兔BMP7成熟蛋白有BMPs特有的7个位置固定的半胱氨酸残基和TGF-β家族指纹。家兔BMP7 3′UTR区转录终止信号的可选择性可能与基因转录后调控有关。
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-β 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.
出处
《遗传》
CAS
CSCD
北大核心
2008年第7期885-892,共8页
Hereditas（Beijing)
基金
河南省教育厅自然基金项目(编号:2007230003)
河南省科技攻关项目(编号:0524030011)资助~~
