Partial analysis of sequences expressed by human osteoblasts {\it in vitro\/} using differential display-PCR (DD-PCR)

Jason Eric Hill, Thomas Jefferson University

Abstract

Bone mineralization is a complex process that has not been fully elucidated. Human osteoblasts isolated from hip explants were cultured with (OB+) and without (OB) beta-glycerol phosphate ($\beta$GP), a promoter of mineralization. Complementary DNA from OB and OB+ cells was amplified by the differential display polymerase chain reaction (DD-PCR). This methodology identified a DD-PCR fragment from mineralized osteoblasts that was absent from non-mineralized cells. This fragment detected two different cDNAs in an OB+ cDNA library screen. One cDNA matched an EST and was expressed in many tissues, including both OB and OB+ RNA, although preliminary data indicated it may be expressed more highly in OB+ than OB cells. The second cDNA matched human mitochondrial DNA. The initial DD-PCR fragment was also cloned directly and yielded one potential recombinant of the correct size. However, the sequence encoded by this clone was not detected in osteoblasts by northern blot analysis. ^

Subject Area

Biology, Molecular|Biology, Genetics

Recommended Citation

Jason Eric Hill, "Partial analysis of sequences expressed by human osteoblasts {\it in vitro\/} using differential display-PCR (DD-PCR)" (January 1, 1996). ETD Collection for Thomas Jefferson University. Paper AAI1381336.
http://jdc.jefferson.edu/dissertations/AAI1381336

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