Molecular characterization of an 18 kb segment of DNA puff C4 of Bradysia hygida (Diptera, Sciaridae)

• Published in: Chromosoma Vol. 103; no. 10; pp. 715 - 724
• Main Authors: Monesi, N, Fernandez, M.A, Fontes, A.M, Basso, L.R. Jr, Nakanishi, Y, Baron, B, Buttin, G, Paco-Larson, M.L
• Format: Journal Article
• Language: English
• Published: Austria 01-07-1995
• Subjects: Amino Acid Sequence; amino acid sequences; animal proteins; Animals; autosomes; Base Sequence; Chromosome Mapping; Chromosomes; Cloning, Molecular; complementary DNA; Diptera; Diptera - genetics; dna amplification; DNA, Complementary; Female; genbank/u13883; genbank/u13892; Gene Amplification; gene expression; genes; Genes, Insect; Insect Proteins; introns; messenger RNA; Molecular Sequence Data; nucleotide sequences; open reading frames; Protein Conformation; salivary glands; Salivary Proteins and Peptides - chemistry; Salivary Proteins and Peptides - genetics; Sciaridae
• ISSN: 0009-5915; 1432-0886
• DOI: 10.1007/BF00344233

The data presented here are an extension of the molecular characterization of DNA puff C4 of Bradysia hygida. A cDNA related to a gene amplified in this puff and expressed when puff C4 expands was cloned and sequenced. Analysis of the amino acid sequence deduced from the open reading frame present in the cDNA indicate that the encoded protein is secreted and comprises mostly alpha-helical coiled-coil. An 18 kb genomic segment containing the transcription unit of this gene was also cloned and the structure and expression of the 1.4 kb mRNA was determined. Quantitative slot blot hybridization of DNA complementary to the transcription unit shows that this gene is amplified about 21 times in the salivary gland, confirming data previously obtained. Fragments upstream of the 5' end, and beyond the 3' end, of the gene transcription unit were also analysed and shown to be amplified at least eight and five times, respectively. Based on these data we discuss how amplification could occur at DNA puffs.