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目的：克隆人骨形态发生蛋白－２（ｈＢＭＰ－２）完整成熟肽基因。方法：依据Ｇｅｎｂａｎｋ中ｈＢＭＰ－２的序列化学合成两条引物，从人胎儿骨组织中提取得到的总ＲＮＡ中，通过反转录聚合酶链式反应（ＲＴ－ＰＣＲ）得到ｈＢＭＰ－２完整成熟肽基因。将所得的基因片段插入克隆载体ｐＵＣ－１９并转化大肠杆菌ＪＭ１０９，提取重组质粒，酶切鉴定并测序。结果：ＤＮＡ琼脂糖凝胶电泳显示：ＰＣＲ产物为一长约４００ｂｐ的带，阳性克隆质粒经双酶切可切出约４００ｂｐ的片段。全自动 ＤＮＡ测序表明与 Ｇｅｎｂａｎｋ中的序列完全相符。结论：通过反转录聚合酶链式反应从人胎儿骨组织中成功地克隆了人ＢＭＰ—２完整成熟肽基因，基因序列完全正确。
Objective: To clone human bone morphogenic protein-2 (hBMP-2) complete mature peptide gene. Methods: Two primers were synthesized according to the sequence of hBMP-2 in Genbank. The complete mature peptide of hBMP-2 was obtained by reverse transcription-polymerase chain reaction (RT-PCR) from total RNA extracted from human fetal bone tissue gene. The resulting gene fragment was inserted into the cloning vector pUC-19 and transformed into E. coli JM109, recombinant plasmid was extracted, identified by restriction analysis and sequencing. Results: DNA agarose gel electrophoresis showed that the PCR product was a band of about 400bp, and the positive cloned plasmid was double digested to cut a fragment of about 400bp. Fully automated DNA sequencing showed complete agreement with the sequences in Genbank. Conclusion: The full-length mature human BMP-2 gene was successfully cloned from human fetal bone tissue by reverse transcription-polymerase chain reaction. The gene sequence was completely correct.