Cloning and characterization of genes that are responsive to various hormones in 3T3-L1 adipocytes

• Main Authors: Cheng-chih Shih, 史承治
• Other Authors: Jim C. Fong
• Format: Others
• Language: zh-TW
• Published: 2004
• Online Access: http://ndltd.ncl.edu.tw/handle/63067095446296836899

碩士 === 國立陽明大學 === 生物化學研究所 === 92 === Abstract Our lab used 3T3-L1 adipocyte as materials to study some novel genes. Ather treating cells with various substances, suppression subtractive hybridization ( SSH) was employed to generate some subtractive cDNA clones. These EST clones were analyzed by bioinformatics. In the progress of finding some novel genes, we chose two genes as our study target, and the functions of the two genes are unknown so far. But analyzing the databases, we found that the products of these two genes have different splicing forms. The purpose of this thesis is to explore the properties and functions of these two genes. In the NCBI database, the accession number of the first gene is AK077556 (long form). We made use of bioinformatics to search some information about this gene, and we got that this gene has the other two splicing forms. The accession number of these two splicing forms is BC012275 (short form) and AK037644 (middle form), respectively. From the protein database, we knew that long form and middle form both have zf-MYND (conserved zinc finger- hydrophobic heptad repeat) and ANK (ankyrin repeats) domains, but short form only has ANK domain. From the result of RT-PCR (Reverse transcription – Polymerase Chain Reaction ), we found that these three splicing forms have different reaction patterns treating with varios substances. The accession number of the second gene is BC009125. Analyzing the sequence of this protein, we knew that the protein has U box (modified RING finger) domain. But the other splicing form of this gene is out of our expectation. Because we didn’t find it in the alternative splicing database. Interestingly, we found it from agarose gel electrophoresis. There always has another band under our target band. From the result of RT-PCR, we found that these two splicing forms also have different patterns treating with various substances. By analyzing the information in databases, we noted that these two genes have homology among species, including homo sapiens. We also found that these two genes maybe involved in some cancer cell lines. So, there is still a long way to go to figure out the functions of these two genes.