恶性疟多抗原表位融合基因在不同载体中的克隆与表达  

Cloning and expression of hybrid antigen of alaria Plasmolium falciparum

作　　者：王衣祥[1] 曹诚[2] 毛盛贤[1] 马清钧[2] 

机构地区：[1]首都师范大学生物系 [2]军事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2001年第3期188-191,共4页Letters in Biotechnology

摘　　要：通过PCR方法扩增并分析了恶性疟原虫多抗原表位融合基因awte序列 ,再分别克隆于温度诱导型融合表达载体pEX31b和IPTG诱导型融合型表达载体pGEMEX1中 ,并进行了诱导表达 ,并对表达产物MS2 AWTE和T7Φ10 AWTE融合蛋白进行间接ELISA检测 ,证实MS2 AWTE和T7Φ10 AWTE都具有免疫学活性。另外 ,同样将awte基因分别克隆于温度诱导型和IPTG诱导型非融合表达载体pBV2 0 0、pKEN中并进行诱导表达 ,但在SDSawte, a multiple-stages and multiple-antigens hybrid epitopes gene of Plasmolium falciparum, was amplified by PCR, sequenced by automated by DNA sequencer and cloned into pBV220, pEX31b, pKEN and pGEMEX1 vector. The recombinant thermal inducible plasmids pBV220-AWTE and pEX31b-AWTE were introduced into E.coli strains. After a heat shock from 30℃ to 42℃, the fusion protein MS2-AWTE was expressed by pEX31b-AWTE engineering bacteria while non-fusion protein AWTE was not expressed by DH5α(pBV220-AWTE). The recombinant IPTG inducible plasmids pKEN-AWTE and pGEMEX1-AWTE were introduced into E.coli strains. After IPTG induction, There was no obivious expression in non-fusion form AWTE by DH5α(pKEN-AWTE), but the gene does expressed pGEMEX1-AWTE engineering bacteria when fused to T7Φ10 at C terminal. Fusion proteins of MS2-AWTE and T7Φ10-AWTE were partially purified and show to be immunogenic by indirect enzyme linked immune adsorbent assay (ELISA).

关 键 词：PCR 基因克隆 表达 融合蛋白 ELISA 恶性疟原虫 多抗原表位融合基因 awte 载体 

分 类 号：R382.31[医药卫生—医学寄生虫学] R394.8[医药卫生—基础医学]