Immune activation upregulates lysozyme gene expression in Aedes aegypti mosquito cell culture

Y. Gao, A. M. Fallon

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

After stimulation with heat-killed bacteria, cultured cells from the mosquito Aedes aegypti (Aag-2 cells) secreted an induced protein with a mass of ≈ 16 kDa that cross-reacted with antibody to chicken egg lysozyme. To investigate whether lysozyme messenger RNA is induced in bacteria-treated cells, we used polymerase chain reaction-based approaches to obtain the complete lysozyme cDNA from Aag-2 cells. The deduced protein contained 148 amino acids, including a 23 amino acid signal sequence. The calculated mass of the precursor protein is 16 965 Da, which is processed to yield a mature lysozyme of 14 471 Da with a calculated pl of 10.1. The lysozyme from Ae. aegypti shared 50% amino acid identity with lysozymes from Anopheles gambiae and Anopheles darlingi, which in turn shared 70% identity between each other. Northern analysis with the lysozyme cDNA probe showed induction of a 1.3 kb messenger RNA during the first 3 h after treatment of Aag-2 cells with heat-killed bacteria; followed by maximal expression 12-36h after treatment. Southern analysis suggested that the gene likely occurs as a single copy in the genome of Aag-2 cells.

Original languageEnglish (US)
Pages (from-to)553-558
Number of pages6
JournalInsect molecular biology
Volume9
Issue number6
DOIs
StatePublished - 2000

Keywords

  • Immunity proteins
  • Insect cell lines
  • Insect immunity
  • Northern blot
  • RT-PCR
  • Southern blot

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