[cDNA clones of differential expression between human normal liver tissue and hepatocarcinoma].
Daqian Wan, Xiaoni Zhang, Xiaolan Zhou
Abstract
Daqian Wan, Xiaoni Zhang, Xiaolan Zhou
Abstract
Using mRNA differential display technique, we studied the interesting cDNA clones of differential expression between human normal liver and hepatocarcinoma. mRNA extracted from normal liver tissue and hepatocarcinoma cell line Hep3B were subjected to RT-PCR reaction with ten combinations of two 3' primers and five 5' primers. After subjecting these PCR products to labelling with 35S(a)-dATP, PAGE and autoradiography, we obtained a lot of cDNA clones of differential expression. Thirty-one of these clones were partially sequenced. It was shown that 30 clones had no sequences matched with GenBank except one. Dot hybridization showed that 2 cDNA clones were overexpressed in liver cancer tissue and 1 cDNA clone was overexpressed in normal liver tissue. The further characterization of these cDNA clones is in progress.
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Using mRNA differential display technique, we studied the interesting cDNA clones of differential expression between human normal liver and hepatocarcinoma. mRNA extracted from normal liver tissue and hepatocarcinoma cell line Hep3B were subjected to RT-PCR reaction with ten combinations of two 3' primers and five 5' primers. After subjecting these PCR products to labelling with 35S(a)-dATP, PAGE and autoradiography, we obtained a lot of cDNA clones of differential expression. Thirty-one of these clones were partially sequenced. It was shown that 30 clones had no sequences matched with GenBank except one. Dot hybridization showed that 2 cDNA clones were overexpressed in liver cancer tissue and 1 cDNA clone was overexpressed in normal liver tissue. The further characterization of these cDNA clones is in progress.
Key concepts: Complementary DNA, Molecular biology, clone (Java method), Biology, GenBank, Differential display, Liver tissue, Messenger RNA