期刊信息
- 刊名： 河北师范大学学报（自然科学版）Journal of Hebei Normal University (Natural Science)
- 主办： 河北师范大学
- ISSN： 1000-5854
- CN： 13-1061/N
- 中国科技核心期刊
- 中国期刊方阵入选期刊
- 中国高校优秀科技期刊
- 华北优秀期刊
- 河北省优秀科技期刊
The Construction and Expression of New-type rPA(K)Prokaryotic Vector
摘要/Abstract
摘要：
利用PCR技术,获得了rPA(K)cDNA片段,将其克隆至pET-30a(+),成功构建了新型的rPA(K)原核载体pLrA(K),并实现了其在大肠杆菌中的初步表达,经IPTG(终浓度为1mmol/L)分别于37℃诱导1,2,3h后,以Bio-Rad凝胶成像仪分析目的蛋白,相对含量分别是19%,15.8%和24.3%,平均密度分别是0.13,0.14和0.16,IPTG诱导3h后蛋白表达量最高.ELISA结果显示表达产物与抗t-PA抗体呈现特异性阳性反应,进一步参考标准曲线得出样品中rPA(K)的平均含量为210μg/L.
Abstract：
By using PCR,the rPA(K)cDNA was got,then it was cloned into pET-30a(+)to get the newtype rPA(K)prokaryotic vector and it was expressed successfully.The initial content of target pro tein went to 24.3% and the average density was 0.16.The molecular weight was about 43 ku.The ELISA indicated the expressive product had specific positive response to anti-t-PA antibody and its average content w as 210 μg/L.
关键词
关键词：
组织型纤溶酶原激活剂
;
rPA(K)
;
原核表达
Key words：
t-PA
;
rPA(K)
;
prokaryotic expression