[Chemical synthesis and cloning of Plasmodium falciparum 45 peptide antigen gene].
We have synthesized a 162 bp gene of human Plasmodium falciparum hybrid peptide antigen by the solid-phase phosphoramidite method with ABI 381A DNA synthesizer. The gene encodes three fragments of the relative molecules 83 kDa, 55 kDa and 35 kDa merozoite-specific proteins and two CS repeats or four peptides. The gene with the designed two cohesive ends was divided into 8 fragments to be synthesized. All synthetic fragments were annealed and ligated with T4 DNA ligase to form double DNA chain. This synthetic gene was recombined with P-Blue script as vector and transformed into E. coli JM109. The positive recombinants were screened out by dot hybridization and enzyme analysis. The DNA sequence analysis showed that the synthesized human Plasmodium falciparum hybrid peptide antigen gene was identical with the designed one. (Figs. 1-4).
Qiu
M Y
MY
Institute of Genetics, Fudan University, Shanghai.
Min
Y J
YJ
Wang
Q S
QS
Zhong
X L
XL
Cheng
S R
SR
Wang
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000138:chemical synthesis / Q000235:genetics; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D017125:Genes, Protozoan; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics*
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D004274:DNA, Recombinant