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Characterization of genomic DNA encoding cecropins from an Aedes albopictus mosquito cell line
Author(s) -
Sun D.,
Fallon A. M.
Publication year - 2002
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.0962-1075.2001.00305.x
Subject(s) - biology , gene , genomic dna , cecropin , microbiology and biotechnology , complementary dna , intergenic region , genetics , coding region , dna , intron , genome , antimicrobial peptides , bacteria
We used cDNA probes from Aedes albopictus mosquito cecropins AalCecA , B , and C to obtain genomic DNA copies and flanking DNA. Two gene copies ( AalCecA1 and A2 , AalCecB1 and B2 , AalCecC1 and C2 ) encoding each of the three mature cecropin peptides were recovered. All these genes had a similar organization, into two exons interrupted by a single short intron. AalCecA1 and AalCecA2 encode mature protein products that differ by one amino acid residue, while AalCecB1 and AalCecB2 , AalCecC1 and AalCecC2 encode identical mature cecropin peptides, respectively. The AalCecB and C gene pairs each share a common intergenic region of approximately 1 kb, with the two coding regions transcribed in opposite directions. With the exception of small insertions/deletions, the intergenic spacer region was highly conserved between the B1/C1 and B2/C2 clones. In transfected cells, 0.8 kb of upstream sequence was sufficient for inducible expression of AalCecA1 . Within this region, a 28 bp sequence at positions –192 to –165 upstream of the transcription initiation site was found to contain a potential regulatory element. In electrophoretic mobility shift assays, synthetic double‐stranded DNA containing this 28 bp sequence retarded protein in cytoplasmic and nuclear extracts from C7‐10 cells.