1. Methods The total RNA was acquired from young rabbit particular cartilage, the BMP7 gene was inserted into pcDNA-3.1 to construct eukaryotic expression vector plasmid of pcDNA3.1-BMP7; the target gene-BMP7 was identified respectively by PCR analysis, restriction enounces analysis and nucleotide sequencing; the plasmid was transfected into adult rabbit knee articular chondrocytes, then the expression of BMP7 was detected by in situ hybridization, PCR and Western blotting.
从体外培养的幼兔膝关节软骨细胞中提取总RNA；按GenBank BMP7基因序列化学合成2条引物，采用RT-PCR方法得到BMP7基因；将BMP7基因片段插入到真核表达载体pcDNA 3.1中，构建pcDNA3.1-BMP7真核表达载体质粒；利用双酶切、PCR及核苷酸序列分析鉴定BMP7目的基因；将重组pcDNA3.1-BMP7真核表达载体质粒转染家兔关节软骨细胞，分别采用原位杂交、PCR、Western blotting法测定BMP-7表达。