The Plasmodium Falciparum Drug-Binding Protein To The Antimalarial Drug Artemisinin

1, To construct a cDNA expression library from erythrocytic Plasmodium falciparum(FCC1). The total RNA has benn obtained by using Triplix Kit ; a modified oligo (dT)primer(CDSâ…¢ PCR primer)was used in singIe-strnaded(ss)DNA synthesis reaction ,the ss DNA was reverse transcriptased selective from total RNA directly ; double-stranded(ds)cDNA was selectively amplifecated by Long-Distance (LD)PCR;the cDNA which more than 200bp was collected and purified by Glass-milk kit after Proteinase K digestion and Sfi digestion;this library was constructed after cDNA was ligated to A TiplEx2 phage particle and was packaged with the packaged extract system in vitro.A higher titer and higher recombinant ratio cDNA library was constructed. The cDNA library is suitable for further study. 2 To obtain the translationally controlled tumor protein(TCTP) gene,the total RNA of Plasmodium falciparum from Hainan(FCC1) was extracted by using Triplix Kit ; a modified oligo (dT)primer(CDSâ…¢ PCR primer) was used in single-stranded(ss) DNA synthesis reaction ,double-stranded(ds)cDNA was selective amplifecated by Long-Distance (LD)PCR;According to the known seqence of Plasmodium yoelii,a pari of primers were designed and synthesized,the TCTP gene of Plasmodium falciparum from Hainan(FCCl) was specifically amplified by PCR technique and then the gene was inserted into the PMD-18-T vector,TCTP gene was cloned by the Xl1-blue. The sequencecing result showed that Plasmodium falciparum from Hainan(FCC1) and Plasmodium yoelii exhibited 85% homology in nucleotide sequence and 88% homology in the amino acid sequence ,comparison with the TCTP gene of Plasmodium falciparum from GenBank, Plasmodium falciparum Hainan(FCC1) and Plasmodium faciparum from GenBank exhibited 98% homologyin nucleotide sequence and 97% homology in amino acid sequence.