期刊文献+

恶性疟原虫FCC1/HN株谷氨酸脱氢酶基因的克隆及序列分析

CLONING AND SEQUENCE ANALYZING OF GDH GENE OF PLASMODIUM FALCIPARUM ISOLATE FCC1/HN
下载PDF
导出
摘要 目的 克隆恶性疟原虫海南株 (FCC1/ HN)谷氨酸脱氢酶 (GDH)基因并测定其序列 ,比较 FCC1/ HN株与国外分离株 GDH基因序列的差异。 方法 根据 GDH基因已知序列设计合成一对引物 ,应用 PCR技术从 FCC1/ HN株基因组 DNA中扩增 GDH基因 ,并将其克隆入 p MD18- T载体。阳性克隆的重组质粒经酶切及 PCR鉴定后 ,用双脱氧链末端终止法进行基因序列测定。应用 DNAstar软件比较不同分离株 GDH基因序列的同源性。 结果  PCR扩增得到特异的 FCC1/ HN株 GDH基因序列。酶切及 PCR鉴定获得了正确的 p T- GDH重组质粒。测序表明 ,恶性疟原虫 FCC1/ HN株 GDH基因全长 132 9bp,编码 44 2个氨基酸。序列分析表明 ,我国恶性疟原虫 FCC1/ HN株与国外的 FCQ2 7、K1株GDH基因编码的氨基酸序列有少量的差异。 结论 克隆了恶性疟原虫 FCC1/ HN株 GDH基因 ;序列测定及同源性分析表明 ,FCC1/ HN株与其它分离株的 Objective To clone and determine the nucleotide sequence of the glutamate dehydrogenase (GDH) gene of Plasmodium falciparum isolate FCC1/HN, and analyze the differences of the sequences of GDH genes among isolate FCC1/HN and other ones worldwide. Methods The GDH gene was amplified by PCR technique from genomic DNA of isolate FCC1/HN. Then it was cloned into pMD18 T vector. The positive clones were screened and identified by agarose gel electrophoresis and endonuclease digestion and PCR technique. The cloned GDH gene was determined by the dideoxy chain termination method. And the DNAstar software was used to find out the differences of the sequences of the GDH genes among different P. falciparum isolates. Results The GDH gene of isolate FCC1/HN was specifically amplified, and the correct recombinant plasmid pT GDH was constructed. The result of sequencing the nucleotides showed that the GDH gene of isolate FCC1/HN was 1 329 base pairs in full length, encoding 442 amino acids. P. falciparum isolates FCC1/HN, FCQ27 and K1 have a few different amino acids deduced from the GDH genes. Conclusion The GDH gene of P. falciparum isolate FCC1/HN was successfully cloned and sequenced. Sequences analysis showed that the GDH genes of isolate FCC1/HN and other ones shared high homology.
出处 《中国寄生虫病防治杂志》 CSCD 2001年第4期244-247,共4页 Chinese Journal of Parasitic Disease Control
基金 中山医科大学"2 1 1"重点学科建设课题基金 (No.981 69) 广东省自然科学基金 (No.980 0 89) 教育部博士点基金 (博教 No.93- 1 86)资助
关键词 恶性疟原虫 谷氨酸脱氢酶 克隆 序列分析 GDH 基因编码 Plasmodium falciparum glutamate dehydrogenase cloning, molecular sequence analysis
  • 相关文献

参考文献9

  • 1[1]Wagner JT,Ludemann H,Farber PM,et al. Glutamate dehydroge-nase, the marker protein of Plasmodium falciparum cloning, ex-pression and characterization of the malaria enzyme [ J]. Eur JBiochem, 1998,258(2) :813~819.
  • 2[2]Yuan P and Stewart TS. Plasmodium falciparum glutamate dehydrogenase (GDH) gene, complete cds [DB]. GenBank accessionnumber AF0986 7 5 , 26-MAR-1999.
  • 3[3]Trager W and Jensen JB. Human malaria parasites in continous cul-ture [J]. Science, 1976,193(4254): 673~675.
  • 4[4]Hunt NH and Stocker R . Oxidative stress and redox status ofmalaria-infected erythrocytes [J]. Blood Cells, 1990,16 (2 - 3 ): 499~526.
  • 5[5]Schirmer RH, Muller JG, Krauth-Siegel RL. Disulfide reductase inhibitors as chemotherapeutic agents: the design of drugs for try-panosomiasis and malaria [J]. Angew Chem Int Ed Engl, 1995,34:141~154.
  • 6[6]Vander-Jagt DL, Intress C, Heidrich JE, et al. Marker enzyme ofPlasmodium falciparum and human erythrocytes as indicators ofparasite purity [J]. J Parasitol,1982,68(6)..1068~1071.
  • 7[7]Roth-E Jr. Plasmodium falciparum carbohydrate metabolism: aconnection between host cell and parasite [J]. Blood cells , 1990,16(2-3)..453~460.
  • 8[8]Vander-Jagt DL,Hunsaker LA,Kibirige M,et al. NADPH-produc-tion by the malarial parasite Plasmodiurn falciparum [J]. Blood,1989,74(1):471~474.
  • 9[9]Kutner S,Baruch D,Ginsburg H,et al. Alterations in membrane per-meability of malaria-infected human erythrocytes are related to thegrowth stage of the parasite [J]. Biochim Biophys Acta , 1982,687(1):113~117.

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部