Construction Of Human Astrocytoma CDNA Library And Cloning Of Human Fatty Acid Binding Protein 7 Gene(FABP7)

Glioma is the most representative tumor of human brain tumors. Recesently, since the development of molecular biology and evaluation of new anti-tumor drug, working on the glioma therapy progressed at a rapid rate. For exploition the pathogeny of genesis and progress of the glioma on a molecular level and discussion the pre-diagnosis of the glioma, we constructed the human astrocytoma cDNA library thus there will be a platform for researchment of glioma tumorigenesis related genes.The human astrocytoma cDNA library was constructed by SMART technic. Total RNA was extracted from three samples of human astrocytoma. After purification of mRNA, the first strand cDNA was synthesized by using M-MLV RTase and SMART primers, then amplified the cDNA by LD-PCR (Long-distance PCR) . We evaluated the quantity of cDNA by using agarose gel electrophoresis, then purified the DNA fragments from the agarose gel and cloned in pMD18-T vector. After conversion of vector into E.coliDH5α, we constructed the astrocytoma cDNA library. This library contains 2×10~5 clones. 90% of them are positive clones, most of insert DNA are about 200-2500 bp. These data show that this cDNA library is of reasonably good qualityThen we cloned the human lipid acid binding protein (FABP7) Gene from the library by using the FABP7 primer which was designed by our lab. The gene we cloned has 100% homology with the sequence obtained from Genebank. Next, we investigated the expression of the FABP7 in 5 glioma samples and the coupled normal tissue samples. Finally we find that when the expression of the FABP7 is different between tumor organization and normal organization.