›› 1990, Vol. 8 ›› Issue (4): 249-252.

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CONSTRUCTION OF A cDNA LIBRARY OF SCHISTOSOMA JAPONICVM

  

  • Received:2017-01-09 Revised:2017-01-09 Online:1990-11-30 Published:2017-01-09

Abstract: The template mRNA was extracted from Schistosoma japonicum. The first strand of cDNA was synthesized by AMV-reverse transcriptase. The second strand cDNA was first digested by RNase H to remove mRNA and was then synthesized by AMV-reverse tran-scriptase, T4-DNA polymerase- Sizing of cDNA was applied on a NACS column to remove small fragments of less than 1 kb. Homopolymeric tailing of vector (PUC18) was done with dGTP and DNA terminal transferase and tailing of the cDNA with dCTP was carried out under the same conditions. After annealing, the plasmids with cDNA were transformed into E. coli MC1061. The efficiency of cloning was about 104/μg mRNA with 30% of the transformants having the inserts of cDNA (Figs. 1-2).