β-NGF在真核细胞蛋白表达系统中的表达及活性鉴定

EXPRESSION AND VITAL IDENTIFICATION OR β-NGF IN PROTEIN EXPRESSION SYSTEM IN EUKARYOTE

目的 :得到较高表达 ,且表达产物具有良好生物学活性的神经生长因子。方法 :将神经胶质源 β -NGF基因克隆于DNA转移载体pBacPAK8中 ,获得重组DNA转移载体pBacPAK -NGF ,与线性化Bm -BacPAK6修饰病毒基因组DNA共转染Bm -N细胞 ,经过体内重组 ,筛选到重组病毒。用重组病毒感染家蚕幼虫 ,经SDS-PAGE检测及利用PC12细胞进行NGF生物活力测定。结果 :感染重组病毒BmNPV -NGF5d的家蚕血淋巴有一条与天然NGF单体分子易相近的蛋白组分 ,且有类似于小鼠 2 .5gNGF的活性。结论 :神经生长因子在家蚕幼虫中得到较高表达 。

Objective: to obtain nerve groath factor(NGF) wfich hae biological vitality by gene expressing Methods: Neuroglia cellogen β-NGF renes were cloned in DNA transfering vertor,then they were transfected Bm-BacPAK6.Recombinated viruses were xelected by recombination.Chinese silkworms were infected with recombinated viruses.β-NGF was measured by SDS-PAGE and its vitality was measured with PC12 cell.Results: Chinese silkworm infectde recombinated virus BmNPV-NGF atfer five days had simIlar vitalityte 2.5g NGF of mouse. Conclusion: Nerve growth fator can be expressed and the product has better biological vitality.