恶性疟原虫FCC1/HN株醛缩酶编码区基因的克隆及表达  

Cloning and Expression of Aldolase Encoding Gene of Plasmodium falciparum FCC1/HN Strain

作　　者：张瑞娟[1] 朱淮民[1] 曹毅[1] 周爱国[1] 郑志强[1] 张青锋[1] 郑徽[1] 

机构地区：[1]第二军医大学病原生物学教研室

出　　处：《中国寄生虫学与寄生虫病杂志》2004年第3期133-135,共3页Chinese Journal of Parasitology and Parasitic Diseases

基　　金："十五"国家科技攻关资助项目 (2 0 0 1BA70 5B0 9)~~

摘　　要：目的　克隆恶性疟原虫海南株 (FCC1/HN)株糖酵解醛缩酶 (ALD)编码区基因。　方法　利用已知ALD基因序列设计一对特异性引物 ,从基因组DNA中用PCR扩增ALD基因 ,将其克隆入pQE 3 0载体 ,阳性克隆经酶切鉴定后测序 ,在此基础上将重组质粒转化大肠埃希菌M15进行表达。　结果　PCR扩增后获得特异性扩增片段 ,测序结果显示我国的恶性疟原虫FCC1/HN株与恶性疟原虫 3D7株ALD基因序列完全相同。重组融合蛋白通过镍 次氮基三乙酸 (Ni NTA)亲和层析及阳离子交换层析进行纯化。　结论　我国的恶性疟原虫FCC1/HN株与文献报道的恶性疟原虫 3D7株ALD编码区基因序列相同 。Objective To clone, sequence, and express the aldolase(ALD)encoding gene of Plasmodium falciparum FCC1/HN strain. Methods The ALD encoding gene was amplified by PCR from genomic DNA of FCC1/HN strain. The positive clones were screened and identified by agarose gel electrophoresis and endonuclease. The recombinant plasmid was transformed into E. coli M15. The fusion protein was expressed by IPTG induction and purified by Ni-NTA affinity chromatography and anion exchange column. Results The ALD gene of P. falciparum was amplified. Analysis of sequencing showed that the ALD gene of P. falciparum was identical with the sequence of other reported isolates. A Mr 41 000 fusion protein was induced by IPTG and was purified by chromatography. Conclusion The ALD gene of P. falciparum FCC1/HN strain was identical to the other reported isolates. ALD fusion protein of P. falciparum was expressed and purified.

关 键 词：恶性疟原虫 FCCL HN 醛缩酶 编码区 基因表达 克隆 PCR引物 

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