Construction of cDNA Library of House Fly (Musca domestic)

Total RNA was separated from the abdomen of house fly,after which had been challenged for 24 hours with E.coli and Staphylococcus aureus.Then,the mR NA was purified from it.After that,ds-cDNA was synthesized by reverse transcriptase from mRNA,and small cDNA which was lower than 400 bp was removed by Sep harose CL-4B spun column.After EocRⅠ/NotⅠAdaptors were added to cDNA,those that werent ligated to cDNA were removed by another Sepharose CL-4B spun column.ThecDNA was inserted into λgtll,the recombined vector packaged in vitro,infected a host strain Y1090.Thus,the cDNA library was co structed.The titer of the newly constructed cDNA library was 3.46×105 pfu/mL,and its recombination rate was 99.6%.The library would provide basis for t he cloning of the antimicrobial peptide genes of house fly.