摘要
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。
To clone integral human bone morphogenetic protein - 2(BMP - 2) mature peptide gene. Methods:. Two primers were chemosynthesized according to the hBMP - 2 sequence reported in Genbank. The hBMP - 2 gene was gained by RT - PCR from the cDNA which was extracted for human fetus bone tissue, and was cloned into vector pUC - 19. The seqence of recombinanted plasmid pUC - 19 - hBMP - 2 was analyzed by sequence analysis,. Results: DNA agarose gel electrophoresis showed that the product of RT - PCR was about 400bp, and double enzyme digestion of the recombinanted plasmid corresponded with it. The result of sequence assay was in agreement with the reported hBMP - 2 sequence in Genbank. Conclusion: The integral human BMP - 2 mature peptide gene was cloned successfully, and totally identified with the sequence of human BMP - 2 in Genbank.
出处
《中国临床解剖学杂志》
CSCD
北大核心
2000年第3期252-253,共2页
Chinese Journal of Clinical Anatomy
