Construction Of Liver And Kidney CDNA Library And Sequence Analysis Of Some Functional Genes Of Ctenopharyngodon Idellus

In this paper, the liver and kidney of grass carps were selected as the experimental materials for the cDNA libraries. The libraries were constructed by oligo(dT) primer directed cloning technology. 510 clones of the cDNA library were sequenced and then they were classified and analyzed basing on bioinformatics methods.First of all, mRNA rich in Poly(A) was purified by Oligotex mRNA Kits after the total RNA was extracted using the method of Trizol from the liver and kidney of the grass carp, then Ds- cDNA was obtained after reverse-transcription and PCR. EcoRI adapters were ligated to the blunt ends and the ends were phosphorylated. Ds-cDNA longer than 500bp with both viscous terminals was collected after Sfi I digestion and transferred after ligation with pBlueseriptl lsK(+)vector, then they were transformated into DH-5a. Finally Positive clones were selected to do PCR detection of bacteria.The library reaches 1.15×10~6 in capacity; the percenyage of recombination is as high as 94%. It can meet our future work.And a total of 510 sequences whose length is between 410-840bp were acquired, including 315 non- repetitive sequences. Among the 315 unigenes, 150 unigenes were functionally known. They play roles in transcription regulation, translation, Ribosomal structure, biosynthesis, cytodieresis, immunity, growth, reproduction , special stimulation, physiological regulation, protein folding, modification and degradation, membrane structure, material transport and cellular signal transduction, metabolism and so on. Then the structure and function of total length genes were analysised.Based on the cDNA library, genes can be screened more conveniently. What's more, full length genes will be separated and the function will be investigated more efficiently.