ST细胞酵母双杂交cDNA文库的构建及鉴定.pdf

2017 44 3 667672 China A nim al H usbandry Vete r inary M e d ic ine doi cnki j ST cD NA orcine parvovirus PPV V P p ST cDNA RNeasy M in Kit ST RNA cDNA SM A RT cDNA ST cDNA CFU mL cDNA bp bp ST ST cDNA Q A Construction and Characterization of the Yeast Two hybrid cDNA Library of ST Cells WANG Xin SHI Da DONG Hui CAO Li an CHEN Jianfei y SHI Hong an Z HANG Xin FENG Li y College of L if e Scicnce Northcast A g ricultural Univ ersity H arbin China Div ision o Sw ine In ectious Discases State K e L aborator o Veterinar B iotechnolo f f y y f y gy H arbin Veterinary Research Institute Chinese A cademy of A g ricultural Sciences H arbin China Abstract T o seek out pro teins interact w ith V P structural protein of porcine parvovirus PPV in ST cell and study the molecular mechanisms of viral inf ection and pathogenicity a ST cDN A yeast hy brid library w as created Firtly the total RN A of ST w as ex tracted using RNeasy M in Kit and double strands cDN A w as sy nthesized by SM A RT technique T hen the ST cDN A library w as successf ully created by homologous recombination in y east the titer w as CFU m L the insert s varied f rom to bp the average of inserted f rag ment s w as about bp and the recombination rate w as T hese datas indicated that the yeast tw o hy brid cDNA library w as successf ully constructed and might b