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KMID : 0578319940040050025
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Molecules and Cells
1994 Volume.4 No. 5 p.25 ~ p.29
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A stable hnRNA Species Confined in HeLa Cell Nucleus
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Kim, Jong Hwa
Choi, yang Do/Byun, Si Myung
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Abstract
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To study the stricture and physiological importances of the nucleus confined hnRNA, eDNA clones for relatively stable and abundant hnRNA have been isolated. HeLa cell hnRNA was purified by immunoaftinity chromatography using monoclonal antibody, 4F4, against the hnRNP proteins. First strand eDNA was synthesized using random hexamer as a primer and a eDNA library was constructed in ],.gill vector. Recombinant phage clones of the hnRNA were isolated from the eDNA library by plaque hybridization with radioactively labeled first strand eDNA of the hnRNA as a probe. One of the positive clones, W16W, hybridizes to the discrete species of 1200-1300 by nuclear non-polyadenylated hnRNA. Insert size of the eDNA clone was 809 bp. According to the Southern blotting analysis the hnRNA transcript. is encoded by a single copy gene. Pulse-chase experiment in the presence of actinomycin D reveals that W16W gene is transcribed by RNA polymerase 11 but the hnRNA transcript is relatively stable and quite abundant.
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KEYWORD
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