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Immune activation upregulates lysozyme gene expression in Aedes aegypti mosquito cell culture
Author(s) -
Gao Y.,
Fallon A. M.
Publication year - 2000
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1046/j.1365-2583.2000.00216.x
Subject(s) - lysozyme , biology , aedes aegypti , complementary dna , microbiology and biotechnology , anopheles gambiae , amino acid , messenger rna , gene expression , gene , biochemistry , immunology , botany , larva , malaria
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 pI 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 12–36 h after treatment. Southern analysis suggested that the gene likely occurs as a single copy in the genome of Aag‐2 cells.