Gene expression profiles analyzed by DNA sequencing of cDNA clones constructed from porcine preadipocytes and adipocytes |
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Authors: | Dong-Mok Lee Prati Bajracharya Eun Jeong Jang Eun Ju Lee Sung Hwa Chae Hong-Seog Park Se Won Kang Yong Seok Lee Ki Ho Lee Jongsoo Chang Chung Soo Chung Inho Choi |
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Institution: | 2. School of Biotechnology, Yeungnam University, Gyeongsan, 712-749, Korea 1. Department of Biotechnology, Yeungnam University, Gyeongsan, 712-749, Korea 3. Research Institute of GnC Bio, Daejeon, 305-150, Korea 4. Bovine Resource Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Korea 5. Department of Parasitology, College of Medicine and UHRC, Inje University, Busan, 621-749, Korea 6. Department of Biochemistry and Molecular Biology, Eulji Medical College, Daejeon, 301-832, Korea 7. Department of Agricultural Sciences, Korea National Open University, Seoul, 110-791, Korea 8. Department of Animal Science, Chungbuk National University, Cheongju, 361-763, Korea
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Abstract: | As a step towards understanding the molecular mechanism of adipogenesis in pigs, preadipocytes purified from the back fat of 1 day-old female piglets were used for in vitro culture. Normalized cDNA libraries were constructed with 1.6×107 and 1.1×107 independent clones from preadipocyte and mature adipocyte mRNAs, respectively. Polymerase chain reaction (PCR) result using primers T3 and T7 (universal primer) confirmed the presence of the insert in the vector. Sequencing of 2,112 randomly selected clones from each cDNA library identified 217 clusters, 1,169 singletons, and 216 contigs in preadipocytes and 231 clusters, 1,100 singletons, and 233 contigs in mature adipocytes. Expressed sequence tag (EST) identified 24 genes with known annotation highly expressed in adipocytes and 21 in preadipocytes by at least four EST number. Among those 45 genes, when analyzed by real time RT-PCR, 76% of the gene showed significant difference between preadipocytes and mature adipocytes. Highly expressed genes in mature adipocytes were related to adipogenesis, extracellular matrix control and oncogenes, whereas cytoskeleton-related genes were down-regulated. An interesting similarity found during gene profile studies indicated a correlation between cancer and adipogenesis. |
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