摘要
目的 :探索EB病毒特异性DNA酶 (EBV DNase)可靠实用的制备方法 ,为建立一种新的鼻咽癌 (NPC)血清学诊断方法———血清EBV DNase抗体检测奠定基础。 方法 :采用TPA +正丁酸联合化学诱导的方法诱导处理P3 HR 1细胞株 4 8h ,超声破碎 ,离心收集上清液即得EBV DNase粗提物。用聚丙烯酰胺凝胶电泳 (SDS PAGE)及放免酶活性检测法测定粗提物中的EBV DNase。 结果 :①在SDS PAGE凝胶上 ,与未经诱导的粗提物相比 ,诱导后的粗提物有多条增强表达的蛋白质带 ,尤以相对分子质量为 5 2 0 0 0的蛋白质更加明显。②用放免法酶活性测定时 ,经诱导的粗提物CPM值及根据公式计算的每毫升酶单位 (U/ml)分别 >170 0 0和 4 0 ,而对照物则≤ 2 0 0 0和4 .0。 结论 :联合化学诱导方法能成功地使EBV DNase在P3 HR 1细胞株中增强表达 ,是一种制备EBV
Objectives:To explore a practical method for the preparation of Epstein Barr virus specific DNase(EBV DNase). Methods: P 3 HR 1 cell line was treated with TPA and n butyrate for 48 hours. The cells were then ultrasonicated, the supernatant was collected after centrifugation as EBV DNase crude extract. SDS polyacrylamide gel electrophoresis(SDS PAGE) and radioimmunologic method were used to determine the EBV DNase in the cellular extract. Results:The results of SDS PAGE showed that the expressions of many kinds of proteins were enhanced, especially those with molecular weight of 52000,in the induced cell extract. The CPM value and enzyme unit per miniliter of induced cellular extract was greater than 17000 and 40 respectively in radioimmunological determination. Those of noninduced ones were less than or equal to 2000 and 4.0. Conclusions:Combined chemical induction can enhance the expression of EBV DNase in P 3 HR 1 cell line and is an effective method for the preparation of EBV DNase.
出处
《医学研究生学报》
CAS
2002年第5期381-384,共4页
Journal of Medical Postgraduates
