Functional study of the cis -elements in the IgH locus using bacterial artificial chromosomes.
Item
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Title
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Functional study of the cis -elements in the IgH locus using bacterial artificial chromosomes.
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Identifier
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AAI3187357
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identifier
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3187357
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Creator
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Zhang, Buyi.
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Contributor
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Adviser: Laurel A. Eckhardt
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Date
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2005
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Language
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English
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Publisher
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City University of New York.
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Subject
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Chemistry, Biochemistry | Biology, Molecular | Biology, Genetics
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Abstract
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Cis-elements play important roles in recombination, transcription, and hypermutation events that take place in the IgH locus. The intronic enhancer, Emu is indispensable for V to DJ recombination. The 3'IgH enhancer pair hs3b and hs4 is required for class switch recombination (CSR). Copious transcription from the IgH locus in Ig secreting cells is dependent on an intact 3'IgH region. In this study, reporter genes with the entire 3'IgH regulatory region were constructed using bacterial artificial chromosomes (BACs). These reporter constructs have the advantages of having all the cis-elements in their natural positions and orientations and preserving the possible higher order DNA structure. We found that the 3'IgH regulatory region could confer transgene transcription in most integration sites in an Ig-secreting cell line, but the expression level was not dependent on the copy number of the transgene. However, Emu together with the 3'IgH regulatory region could enable the transgene to be expressed in a copy number dependent manner, suggesting that all the elements in this particular BAC constitute an LCR. In addition, we showed that hs4 deletion from the 3'Igh regulatory region did not affect the ability of this region to confer transgene high level transcription in the same integration sites, suggesting that there is more redundancy in function than previously appreciated in the IgH locus and that hs4 is dispensable for maintaining IgH gene transcription in Ig-secreting cells, even in the absence of Emu.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
