摘要
人乳头瘤病毒(HPV)含有2个晚期开放读码框(ORF),L_2ORF编码的蛋白具有型特异性。本研究用能表达产生完整HPV6bL_2蛋白的质粒p6bL_2NX,采取核酸外切酶Ⅲ定向连续次级克隆的方法,制备了一系列一端逐渐删切的DNA片段,诱生一组从C端至N瑞依次减少的L_2蛋白与相应血清作免疫印迹实验,最小的仍保持阳性反应的多肽即含有抗原表位,实验结果HPV6bL_2蛋白的核酸编码区位于5481-5506之间,其相应氨基酸序列是EPGINPTQ。
PV contains two late open reading frames. T he L_2 protein encoded by L_2 ORF possesestype-specific antigenicity. To map the immunoreactive epitope of HPV 6b L_2,a L_2 expressionplasmid p6bL_2 NX was used.Plasmid p6b L_2 NXK was made by adding K pnI linker intopolylinker of p6bL_2 NXK. Unidirectional 3’ to 5’deletions were made in the HPV 6b L_2 ORFinsert of p6b L_2 NXK by using exonuclease Ⅲ and nuclease S_1.The deleted plasmids ex-pressed a set of carboxy to amino terminus truncated fusion proteins. Region containing theimmunoreactive epitope was mapped by determing retained reactivity of the deleted fusionproteins with sera in Western immunoblot assays. The related plasmids were sequuenced.The immunoreactive epiutope was located at 5481-5506 nts region,and the correspondingpepude was EPGINPTQ.
出处
《中国病毒学》
CSCD
1995年第3期228-231,共4页
Virologica Sinica