Report Abstract

Abstract:

An unknown segment of DNA was recovered from Bactrocera dorsalis and subjected to characterization using insertion into vectors (EMBL3 and pUC19), restriction enzyme mapping, Southern blot sequence verification, and PCR amplification. The genomic library was created successfully with 45 plaques available for analysis. The DNA was positively inserted into the pUC19 vector, as verified by the presence of white colonies on the selective media (LB-amp-Xgal). A restriction enzyme map illustrated that the insert contained recognition sites for HindIII 500 bp, Xho1 1000 bp, and Pst1 3500 bp from the beginning of the insert. The Southern blot verified the presence of the target sequence in the linearized recombinant plasmid. The 720-bp sequence was also analyzed using database analysis, and primers were designed for the amplification reaction. PCR revealed a fragment size of between 100 and 200 base pairs, indicating that the primers successfully isolated the 161-bp target sequence. Following these tests, the unknown segment was found to contain an actin-coding gene. Actin is highly conserved among species. It functions in the eukaryotic cytoskeleton and in muscular contraction.