重组大鼠骨形成蛋白-2成熟肽编码区cDNA克隆

Cloning of Mature Region cDNA of Rat Bone Morphogenetic Protein 2.

目的 :克隆大鼠骨形成蛋白 - 2成熟肽编码区基因。方法 :由大鼠成骨肉瘤细胞株 (UMR10 6)中提取细胞总RNA ,利用逆转录PCR方法 ,从mRNA中扩增出大鼠骨形成蛋白 - 2成熟肽编码区基因cDNA片段 ;将获得基因片段插入pGEM -T质粒中 ,转化到大肠杆菌JM10 9后挑选阳性克隆 ,利用限制性酶切和核苷酸序列分析鉴定重组质粒。结果 :通过质粒DNA酶切分析及序列测定 ,我们获得的基因片段为大鼠BMP - 2成熟区cDNA序列。结论 :克隆获得大鼠骨形成蛋白 - 2成熟肽编码区基因 ,为表达骨形成蛋白 -

Objective: The purpose of this study is to clone rat BMP-2 cDNA that encodes mature rat BMP-2 protein. Methods: Using rat osteosarcoma cell line UMR106 to extract cell total RNA, rat BMP-2 cDNA flanking BMP-2 mature region was amplified by RT-PCR. The rat BMP-2 cDNA was then inserted into cloning vector pGEM-T. Screening by white and blue clone after transforming the recombinant plasmid into E.coli. strain JM109, we got the recombinant plasmid pGEM-BMP-2 which was confirmed by restriction enzyme and sequencing analysis. Results: Restriction enzyme assay and sequencing showed that the construct we got was rat BMP-2 mature region cDNA. Conclusion: We obtained rat BMP-2 mature region cDNA construct which provide us useful target to further expression study. [