Construction and Identification of a cDNA Expression Library from Clonochis sinensis Metacercaria

The cDNA expression library of Clonochis sinensis metacercaria was constructed for selecting stage specific diagnosis antigen gene or vaccine candidate gene.Total RNA of leucocytes was then extracted,pooled together and mRNA was isolated with a column of oligo(dT) cellulose.And cDNA was synthesized by Stratagene cDNA library protocol.The first and second strand of cDNA were synthesized using Stratagene Uni-ZAP XR cDNA synthesis kit,then ligated to EcoR I adapters and digested with Xho Ⅰ.After size fractionated with CHROMA Spin-400 column cDNA were ligated into Uni-ZAP vector and packaged in vitro to construct cDNA libraries.It was found that the size of the primary library was 9.15×105 PFU with a recombination rate of 99.5% and the titer of the amplified library was 1.6×1010 PFU/mL.cDNA library for Clonochis sinensis Metacercaria has been constructed.