期刊
INSECT MOLECULAR BIOLOGY
卷 9, 期 6, 页码 553-558出版社
WILEY
DOI: 10.1046/j.1365-2583.2000.00216.x
关键词
insect immunity; insect cell lines; immunity proteins; RT-PCR; Northern blot; Southern blot
资金
- NIAID NIH HHS [AI 36 258] Funding Source: Medline
After stimulation with heat-killed bacteria, cultured cells from the mosquito Aedes aegypti (Aag-2 cells) secreted an induced protein with a mass of approximate to 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.
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