Construction of cDNA Libraries from Mature Female and Male Worm of Schistosoma japonicum Chinese Main Land Strain

Total RNA of mature female and male worm of Schistosoma japonicum(Sj) were extracted by Trizol.mRNA were further purified through oligo\|dT cellulose.The first strand cDNA was synthesized by using AMV reverse transcriptase.After the synthesis of the second strand,the fragments longer than 500bp were collected by electrophoresis and purified by kit,and were then ligated with lambda ZipLox vector.After package in vitro,two cDNA libraries were constructed, a female cDNA library containing 3 74×10 6 colones,and a male cDNA library containing 3 28×10 6 clones,in which more than 96% clones are recombinant and most of insert DNA were 1kb,more or less.By using PCR we obtained the cDNAs of EGP,23KD membrane protein,Actin and GCP from our cDNA libraries,GCP is a low\|abundant expressed gene.These datas show that two cDNA libraries are of reasonably good quality,and can be used to study the gene differences between female and male worm and to screen the protective antigen genes.