摘要
将人尿激原ｃＤＮＡ分别插入家蚕核型多角体病毒转移载体ｐＢＫ２８３和ｐＢＦ４中，构建成两个重组质粒。所构建的这两个重组质粒与野生型家蚕核型多角体病毒ＤＮＡ共转染家蚕培养细胞，经病毒斑实验筛选出含ｐｒｏ－ＵＫ ｃＤＮＡ的稳定重组病毒株ＢｍＮＰＶ－ｐｋ１和ＢｍＮＰＶ－ｐｋ２。将此两株蛋白平板溶圈测活法和Ｗｅｓｔｅｒｎ印迹法分析细胞培养上清及细胞和家蚕的体液及组织，证实均有ｐｒｏ－ＵＫ表达。
Human pro-UK cDNA was inserted into Bombyx mori nuclear polyhedrosis virus (BmNPV) transfer vectors pBK283 and pBF4. The resulting plasmids and the BmNPV genomic DNA were co-transfected into Bm-N cells, and two stable recombinant viruses BmNPV-pk1 and BmNPV-pk2 were isolated by plaque assay on Bm-N cells. When Bm-N cells and silkworm larvae were infected with recombinant viruses BmNPV-pkl and BmNPV-pk2, the samples of cells fluid and silkworm larvae haemolymph were characterized with Western blot analysis, the blot displayed a major band at 50kDa. Enzymatic acticity in fibrin plate assay was 120IU per mL of cell supernatant. The expression level of pro-uk in the cells fluid 4 days post-infection was 3.1μg/mL, and in infected larvae haemolymph was 30.0μg/mL determined by ELISA.
出处
《北京大学学报（自然科学版）》
CSCD
北大核心
1993年第6期648-654,共7页
Acta Scientiarum Naturalium Universitatis Pekinensis
基金
国家863高科技项目
