摘要
研究了骨形态发生蛋白ＢＭＰ－２ｃＤＮＡ在ＣＯＳ细胞和小鼠肌肉中的表达的情况，从ｐＳＰＳ６５ＢＭＰ－２质粒中回收ＢＭＰ－２ｃＤＮＡ，删除５＇端的非翻译序列，插入ｐＳＶＬ载体中，构建了含有ＢＭＰ－２全长编码序列的重组表达质粒ｐＳＶＬＢＭＰ－２。将表达质粒导入ＣＯＳ－７细胞中，细胞ＲＮＡ点杂交结果表明，转染ＢＭＰ－２基因的细胞内ＢＭＰ－２的ｍＲＮＡ水平明显升高；细胞培养上清的ＥＬＩＳＡ显示，转染ＢＭＰ－２ｃＤＮＡ后，细胞分泌产生的ＢＭＰ－２显著增加。小鼠实验发现，在肌肉内用注射法导入ＢＭＰ－２重组质粒后，局部组织内ＢＭＰ－２的ｍＲＮＡ转录水平也明显提高。
The expression of human BMP-2 cDNA in COS-7 cells and in mice muscles was studied. The gene was obtained from 1578 bp cDNA of BMP-2 by cutting off most of 5' UTR, and inserted into the MCS of eukaryotic expression vector, pSVL. The recombinant ex pression plasmid──pSVLBMP-2 included whole gene sequence encoding 396 amino acids of BMP-2. The pSVLBMP-2 was transfected into COS-7 cell for transient expression through the medium of DEAE-dextran. The RNA dot blot showed that there was transcription of BMP-2 mRNA in the control COS-7 cell at low level, which was obviously enhanced with the pSVLBMP-2 transfected. The immunoreactive assay of ELISA demonstrated the expression of the recombinant protein in the medium from COS-7 cell transfected with pSVLBMP-2. It was also observed that the BMP-2 expressed in mouse muscle by injection of gene preparation directly into the muscle. The fact was found that the injected pSVLBMP-2 could be transcripted in the muscle, its transcript level of BMP-2 mRNA was higher than that of the control. The results provided evidence to indicate the possibility of gene treatment for some specific bone injuries by local gene injection.
