Construction and analysis of subtracted cDNA library by suppression subtractive hybridization from Hyriopsis cumingii liver.

This study used suppression subtractive hybridization(SSH) for construction of the cDNA subtractive library of the liver from the living body infected Hyriopsis cumingii plague virus.After testing,differentially expressed genes were enriched in the 2 10 times that the proof of the cDNA library has strong subtractive efficiency.Three hundred positive clones were randomly picked and identified by PCR method;95% clones contained 0.2-1.0 kb by inserts,which might be the cDNA frag-menu of differentially expressed genes in Hyriopsis cumingii plague virus infected group.Three hundred clones were sequenced and obtained 214 known function EST for 58 different genes were reported in H.cumingii for the first time in this study.According to the classification of functional genes by Adam,the identified EST in H.cumingii fell into eight categories relevant to 2 belong to cell division genes,9 belong to cellular structure and movement genes,10 belong to metabolism genes,7 belong to signal transduction genes,10 belong to cell immune-action genes,20 belong to gene and protein expression associates genes and 26 belong to other proteins with unknown functions.In addition,there are 14 no similar sequence in Genbank,conjecturing its new genes.Results show,the method to construct cDNA subtractive library could well reflect the gene information of abalone affected by H.cumingii plague virus.Results of the present study provide the basic data to research of relationship between resistance breeding and gene expression of liver.