Studies The Difference Expression Of Antibacterial Peptide Gene Lucifensin In Every Growth Phase

Objective:Quantitative PCR, relative quantification assay by Lusifensin defensin gene in Lucilia fly eggs, larvae, pupae, adult expression levels of four to determine the defensin gene expression levels of the strongest periods of Lucilia flies defensin further research and application provide a theoretical basis.Method:1Firstly, corruption spawning fish lure flies will fly species suspected to capture and captive breeding; then fly species obtained from a single community to identify morphological and molecular identification of:(1) morphological identification, in anatomy morphological characteristics of fly species collected under the microscope, with the morphological characteristics of the fly Lucilia comparison;(2) the molecular identification, genomic DNA was extracted fly species, amplified its iTS-1gene fragments and gene fragments CAD, electrophoresis, purification, recovery and sequencing, the sequencing results for homology.2Extraction sericata egg, larva, pupa stage, the adult stage of total RNA by reverse transcription, amplification, cloning and sequencing technology has been part of the sequence of defensin gene Lucifensin will be conservative sequencing results of contrast. According to sequencing results fluorescent quantitative PCR primer design, the use of quantitative PCR relative quantification method with18S rRNA as a reference gene, comparing the growth of Lucilia flies four times defensin expression activity.Results:1Description morphological characteristics and Lucilia flies collected under the microscope is very consistent. ITS1-F/ITS1-R to be as primers, expected product size of360bp, its sequencing results with Lucilia ITS-1gene (accession number:FJ614858.1) similarity of98%; with CAD-F/CAD-R is a primer for amplification, the size of the product expected272bp, and the sequencing results of Lucilia sericata CAD gene (accession number:FJ169332.1) similarity of97%, and only a highly similar DNA sequences. In summary, to determine the species of flies fly Lucilia sericata Lusifensin2gene-based design primers, expected product size of254bp, its sequencing results with Lucilia flies Lusifensin genes.(Accession number:HM243535.1) similarity89%.3defensin expression activity of the larvae of Lucilia flies Lusifensin gene is much higher than the other three times, followed by the adult period, defensin gene expression levels in the order of:larva> adult period> pupae period> eggs period.Conclusion:This trial was the first cloned and sequenced China’s Hunan province sericata ITS-1gene, CAD gene and Lusifensin defensin gene, and conducted a homology. And applying quantitative PCR Lusifensin defensin antimicrobial peptides compared gene expression differences in the activity of the various growth stages of Lucilia flies for the further development of antimicrobial peptides studies provide new ideas.