论文部分内容阅读
目的测定恶性疟原虫FCC1/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PCB技术从FCC1/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JM109感受态细胞,铺x-gal LB平板;挑取阳性茵落,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FCC1/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FCC1/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FCC1/HN株exp-1基因,并测定了其核苷酸序列,为进一步研究其功能奠定基础.“,”Objective To determine the sequence of exported protein 1 (exp-1 )gene of Plasmodium rnfalciparum ( P. F ) isolate FCC1/HN. Methods According to the known sequence of a pair of primers were rndesigned and synthesized. The exp-1 gene was amplified by polymerase chain reaction (PCR)from genomic rnDNA of isolate FCC1/HN and cloned into the pMD-18T vector. Then the recombinant plasmid pT-exp-1 was rntransformed into E. Coli JM109. The positive clones were screened and identified by agarose gel electrophoresis ,rnendonuclease digestion and PCR technique. The correct recombinant plasmid pT-exp-1 was used as template ,rnand the nucleotide sequence of exp-1 gene was determined by the dideoxy chain termination method. Results rnThe exp-I gene from genomic DNA of P. Falcipamm isolate FCC1/HN was specifically amplified, and the rncorrect recombinant plasmid pT-exp-1 was constructed .The full length of exp-1 gene of isolate FCC1/HN,rnencoding 162 amino acids , is 937 base pairs . Conclusion The exp-1 gene of P. Falciparum was cloned and rnsequenced , and a foundation was laid for further study on the function of exp-1 gene.