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目的:克隆人骨形成蛋白2(hBMP2)基因全长,用于临床难治性骨折和骨缺损的再生修复。方法:以成骨肉瘤细胞总RNA为模板,应用反转录-聚合酶链反应法(RT-PCR)克隆hBMP2的cDNA全长;将获得的基因插入pGEM-T-Easy载体质粒,并转化至大肠杆菌后挑选阳性克隆,利用限制性内切酶酶切分析鉴定重组质粒。结果:通过质粒酶切分析和序列测定,获得的基因为hBMP2全长DNA序列。结论:克隆获得hBMP2的基因,为其进一步开发利用提供了前提条件。
OBJECTIVE: To clone the full length of human bone morphogenic protein 2 (hBMP2) gene for clinical refractory fracture and bone defect regeneration and repair. METHODS: Total RNA of osteogenic sarcoma cells was used as a template to clone full-length cDNA of hBMP2 by reverse transcription-polymerase chain reaction (RT-PCR). The obtained gene was inserted into pGEM-T-Easy vector and transformed into After positive clones were selected, recombinant plasmids were identified by restriction enzyme digestion analysis. Results: The full-length hBMP2 DNA sequence was obtained by plasmid restriction analysis and sequencing. Conclusion: The hBMP2 gene was cloned and provided the precondition for its further development and utilization.