Identification of promoter sites in Babesia equi ema-l 5' intergenic nucleotide: I. PCR amplification and restriction mapping

Babesia equi ema-l 5' intergenic 뉴클레오타이드의 프로모터 위치 확인: I. PCR 증폭 및 제한효소지도

  • 곽동미 (워싱턴주립대학교 수의과대학)
  • Published : 2004.03.01

Abstract

Babesia equi ema-1 5' intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5' IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes (Bgll , HindⅢ, Kpn1 and BamH1) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.

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References

  1. Knowles DP, Kappmeyer LS, Perryman LE. 1997. Genetic and biochemical analysis of erythrocyte-stage surfaceantigens belonging to a family of highly conserved proteins of Babesia equi and Theileria species. Mol Biochem Parasitol 90 : 69-79
  2. Kumar S, Yokoyama N, Kim JY, et al. 2004. Expression of Babesia equi EMA-1and EMA-2 during merozoite developmental stages in erythrocyte and their interaction with erythrocytic membrane skeleton. Mol Biochem Parasitol 133 : 221-227 https://doi.org/10.1016/j.molbiopara.2003.10.010
  3. Kappmeyer LS, Perryman LE, Knowles DP Jr. 1993. A Babesia equi gene encodes a surface protein with homology to Theileria species. Mol Biochem Parasitol 62 : 121-124
  4. Knowles DP Jr, Kappmeyer LS, Stiller D, et al. 1992. Antibody to a recombinant merozoite protein epitope identifies horses infected with Babesia eoui. J Clin Microbiol 30 : 3122-3126
  5. Knowles DP Jr, Perryman LE, Goff WL, et al. 1991. A monoclonal antibody defines a geographically conserved surface protein epitope of Babesia equi merozoites. Infect Immun 59 : 2412-2417
  6. de Koning-Ward TF, Speranca MA, Waters AP, et al. 1999. Analysis of stage specificity of promoters in Plasmodium berghei using luciferase as a reporter. Mol Biochem Parasitol 100 : 141-146
  7. de Koning-Ward TF, Fidock DA, Thathy V, et al. 2000. The selectable marker human dihydrofolate reductase enables sequential genetic manipulation of the Plasmodium berghei genome. Mol Biochem Parasitol, 106 ; 199-212
  8. Cowman AF, Baldi DL, Healer J, et al. 2000. Functional analysis of proteins involved in Plasmodium falciparum merozoite invasion of red blood cells. FEBS Lett 476 : 84-88
  9. de Koning-Ward TF, Janse CJ, Waters AP. 2000. The development of genetic tools for dissecting the biology of malaria parasites. Annu Rev Microbiol 54 : 157-85