家蝇cDNA文库的构建  

Construction of cDNA Library of House Fly (Musca domestic )

作　　者：王来元[1] 王金星[1] 赵小凡[1] 王来城[1] 康翠杰[1] 

机构地区：[1]山东大学生命科学学院生物系,济南250100

出　　处：《Zoological Research》2001年第2期159-162,共4页动物学研究（英文）

基　　金：山东省自然科学基金资助项目 !(Y97D0 10 6 0 )

摘　　要：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 mRNA 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 Sepharose CL 4B spun column. After Eoc RⅠ/ Not Ⅰ Adaptors were added to cDNA,those that weren’t ligated to cDNA were removed by another Sepharose CL 4B spun column. The cDNA was inserted into λgtll,the recombined vector packaged in vitro ,infected a host strain Y1090. Thus,the cDNA library was constructed. The titer of the newly constructed cDNA library was 3 46×10 5 pfu/mL,and its recombination rate was 99 6%. The library would provide basis for the cloning of the antimicrobial peptide genes of house fly.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 mRNA 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 Sepharose CL 4B spun column. After Eoc RⅠ/ Not Ⅰ Adaptors were added to cDNA,those that weren't ligated to cDNA were removed by another Sepharose CL 4B spun column. The cDNA was inserted into λgtll,the recombined vector packaged in vitro ,infected a host strain Y1090. Thus,the cDNA library was constructed. The titer of the newly constructed cDNA library was 3 46×10 5 pfu/mL,and its recombination rate was 99 6%. The library would provide basis for the cloning of the antimicrobial peptide genes of house fly.

关 键 词：家蝇 CDNA文库 重组率 文库滴度 抗菌肽 

分 类 号：Q969.538[生物学—昆虫学]