Objective To screen differential expression genes between normal liver and hepatic fibrosis in rats and to demonstrate the molecular mechanism from gene.
Methods mRNA differential display PCR (DD-PCR) was used. First both total RNAs were isolated and reversely transcribed into cDNAs respectively. The products of PCR were run on 6% denaturing polyacrylamide gels(PAG)and autoradiographs were examined to find the differential bands between both gels. The differential bands were reamplified using the same PCR condition as before. Reverse northern analysis was used to identify the bands. After sequencing, the DNA sequence was checked with GenBank data for homology.
Results 16 differential bands were selected from the gel and 12 bands were special by repeated PCR. In the 4 sub-clones, 2 differentially expressed fragments were obtained(N2, F1)by reverse northern hybridization. Homology comparing, the sequence of N2 had 94% homology with Mus musculus apolipoprotein a5(apoa5), but F1 was not found.
Conclusion By mRNA differential display, mRNAs from normal liver and hepatic fibrosis were compared to find one differentially expressed liver-fibrosis-related gene fragment. It will be effective to study the gene display to find the special curative method.
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