摘要
以HBV全基因组质粒为模板,扩增出目的区段:S(1-222AA),preS1(10-50AA)和C(2-30AA),以酶切-PCR的方法进行连接,克隆入表达载体pPIC3.5k,电穿孔转化后利用G418加压筛选出多拷贝整合菌株,诱导表达并对表达产物进行检测。结果显示融合后的基因为ss1c,长度约为906bp。在酵母中成功地进行了表达,表达产物(SSIC)的大小约为31kDa,并形成直径约为22nm的病毒样颗粒。表达产物与HBs抗体、preS1抗体和HBc抗体均有特异反应。为进一步研究治疗性疫苗提供了重要的基础。
PCR method was used to Construct multi-epitope gene sslc containing HBV S （1-222AA）, preS1（10-50AA）and C（1-30AA） gene. The fusion sslc gene was cloned into the expression vector pPIC3.5k. The recombinant vector was transformed into the host cell of GS115 with electroporation method. After screening with G418 and inducing with methanol, the product was analyzed with Western blot and EIA. The fusion gene （SSIC） with the length of 906bp was successfully expressed in yeast system. The molecular weight of SSIC expressed protein was about 31kDa and virus-like particles with diameter of about 22nm were formed. The product had specific reaction with anti-HBs, anti-preS1 and anti-HBc mAbs. Whether SSIC is a promising candidate for therapeutic vaccine for control of chronic HBV infection needs to be further studied.
出处
《中国病毒学》
CSCD
2005年第5期485-489,共5页
Virologica Sinica
