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[Screening and identification for cDNA of differentially expressed genes in human primary hepatocellular carcinoma]
Jing Li1, Ben-li Han, Gui-jun Huang
1Department of Hepatobiliary Surgery of Xinqiao Hospital, Third Military Medical University, Chongqing, PR China. xqyylijing@163.net
Objective:
Screening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC).
Methods:
The differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis.
Results:
Thirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC.
Conclusion:
The subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC.