Sequence determination of exp- 1 Gene of Plasmodium Falciparum Isolate FCC1／HN

目的 测定恶性疟原虫FCCl／HN株exp-1基因序列。方法 根据exp-1基因已知序列设计合成1对引物，用PCR技术从FCCl／HN株基因组DNA中扩增exp-1基因；将exp-1基因克隆入pMD-18T载体，转化大肠杆菌JMl09感受态细胞，铺x—gal LB平板；挑取阳性菌落，用酶切，PCR扩增进行鉴定。以正确的重组质粒为模板，用双脱氧链末端终止法测定exp-1基因序列。结果 从恶性疟原虫FCCl／HN株基因组DNA中获取exp-1基因，成功克隆入pMD-18T载体；测序表明FCCl／HN株exp-1基因全长937bp，编码162个氨基酸。结论 克隆了恶性疟原虫FCCl／HN株exp-1基因，并测定了其核苷酸序列，为进-步研究其功能奠定基础。

Sequence determinationof exp-1 Gene of Plasmodium Falciparum Isolate FCC1/HN

Objective To determine the sequence of exported protein 1(exp-1) gene of Plasmodium falciparum(P. f )isolate FCC 1/HN. Methods According to the known sequence of a pair of primers were designed and synthesized. The exp-1 gene was amplified by polyrnerase chain reaction(PCR) from genomic DNA of isolate FCC I /HN and cloned into the pMD1-18T vector. Then the recombinant plasmid pT-exp-1 was transformed into E. coli JM109. The positive clones were screened and identified by agarose gel electrophoresis endonuclease digestion and PCR technique. The correct recombinant plasmid pT-exp-1 was used as template and the nucleotide sequence of exp-1 gene was determined by the dideoxy chain termination method. Results The exp-gene from genomic DNA of P. falciparum isolate FCC l/HN was specifically amplified, and the correct recombinant plasmid pT-exp-1 was constructed . The full length of exp-1 gene of isolate FCC1/HN, encoding 162 amino acids is 937 base pairs . Conclusion The exp-1 gene of P. falciparum was cloned and sequenced , and a foundation was laid for further study on the function of exp-1 gene.