昆虫杆状病毒表达系统中人骨形成蛋白3的表达及鉴定

探索人骨形成蛋白在昆虫杆状病毒系统中的表达规律。方法将ｈＢＭＰ３全长ｃＤＮＡ克隆入转移载体ＢａｃＰＫ８中，酶切鉴定后，将此载体与病毒ＤＮＡ经脂质体包裹转染昆虫细胞，重组病毒经蓝白选后，提取其ＤＮＡ进行ＰＣＲ反应，鉴定目的基因。收集重组病毒感染５ｄ的细胞进行免疫荧光及电子显微镜观察。结果克隆了目的基因的重组载体经内切酶消化后，琼脂糖电泳示有相应大小的目的基因片段切下，ＰＣＲ电泳结果见有目的基因片段的

Expression and characterization of human bone morphogenetic protein 3 in baculovirus expression syste

AIM To study the expression of human bone morphogenetic protein (hBMP3) in the baculovirus system. METHODS cDNA encoding human bone morphogenetic protein 3 was cloned into the transfer vector BacPK8. The recombinant vector harboring hBMP3 cDNA was identified by restriction enzymes. The linearized virus DNA and the recombinant vector were mixed with lipofectin then transfected into Sf9 cells. The recombinant virus was identified by LacZ blue white selection, PCR analysis, immunofluorescence and electromicroscopic analysis were also made. RESULTS The target DNA fragment in agarose electrophoresis was seen when recombinant transfer vector was incubated with restriction enzyme. The amplified fragment of target gene from recombinant virus DNA by PCR was observed. The positive immunofluorescence granules were observed in cytoplasm and cytomembrane. Unlike wild virus, no polyhedrosis body formation was seen in the recombinant virus under electromicroscopy. CONCLUSION hBMP3 has been expressed in this baculovirus expression system.