Expression of the Plasmodium knowlesi circumsporozoite antigen in Escherichia coli directed by Plasmodium bacterial-like promoter sequences

Ariel Ruiz i Altaba, Luiz S. Ozaki, Fidel Zavala, G. Nigel Godson

Research output: Contribution to journalArticle

Abstract

The Plasmodium knowlesi circumsporozoite (CS) gene is expressed in Escherichia coli directly from a parasite genomic DNA fragment, using promoter and ribosome-binding site (RBS) sequences present in this fragment. Transcription of the CS gene in E. coli is directed by tandem Plasmodium bacterial-like promoter elements located within the 0.5-kbEcoRI-HindIII fragment roughly 2.5 kb 5' from the CS gene within the 11-kb EcoRI parasite genomic DNA fragment. No readthrough from vector promoters or fortuitous promotion from plasmodial A + T-rich sequences was observed. The endogenous Plasmodium promoter of the CS gene does not seem to be recognized by E. coli RNA polymerases. Two tandem E. coli-recognized promoters are relatively strong judging by their ability to drive the bacterial chloramphenicol acetyl-transferase (CAT) gene. Translation of the message must be achieved by utilising an AAGAA sequence 4 bp 5′ from the ATG initiation codon as RBS.

Original languageEnglish (US)
Pages (from-to)135-144
Number of pages10
JournalGene
Volume41
Issue number2-3
DOIs
StatePublished - 1986
Externally publishedYes

Keywords

  • CS gene
  • Recombinant DNA
  • antibiotic resistance
  • malaria parasite

ASJC Scopus subject areas

  • Genetics

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