The isolation and characterization of a P. Angulosus homeobox

 

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dc.contributor.advisor Von Holt, Claus en_ZA
dc.contributor.author Pfeffer, Peter Lance en_ZA
dc.date.accessioned 2016-09-28T19:05:27Z
dc.date.available 2016-09-28T19:05:27Z
dc.date.issued 1991 en_ZA
dc.identifier.citation Pfeffer, P. 1991. The isolation and characterization of a P. Angulosus homeobox. University of Cape Town. en_ZA
dc.identifier.uri http://hdl.handle.net/11427/21983
dc.description Bibliography: pages 93-108. en_ZA
dc.description.abstract The aim of this thesis was to isolate and characterize a homeobox-containing gene of the South African sea urchin Parechinus angulosus. This was achieved by constructing a genomic library of several individuals and screening this library using a probe containing the Antennapedia homeobox. Eight clones were isolated and shown to represent different alleles of the same gene. One clone was sequenced, revealing a homeobox which was termed PaHboxl. This homeobox was compared to published homeobox sequences and shown to be a member of the Antp (Hoxl.l) subclass (table 1.1). A splice donor site was identified 23 bp upstream of the homeobox and the observation confirmed by RNAase mapping. PaHboxl is situated in a genomic area showing a significantly higher degree of restriction fragment polymorphism than expected. This was shown by a statistical analysis which should be of general value in the interpretation of such polymorphisms. The expression of PaHboxl was examined by RNAase protection assays and Northern blotting. Two distinct phases of expression were observed - during embryogenesis PaHboxl is expressed transiently at low levels in 11,5 hr mesenchyme blastula stage embryos (44 ± 8 transcripts per embryo) with levels 3-5 fold lower 2,5 hr before and after this stage. Expression is observed again at up to 160 fold higher levels in the adult with maximal expression in testis (11 transcripts per 10 pg total RNA), and increasingly lower levels in intestines, ovary and Aristotle's lantern. Two transcripts of size 5,2 and 5,7 kbp were observed. Expression in Aristotle's lantern and embryonic stages could not be detected by Northern analysis. en_ZA
dc.language.iso eng en_ZA
dc.subject.other Biochemistry en_ZA
dc.title The isolation and characterization of a P. Angulosus homeobox en_ZA
dc.type Doctoral Thesis
uct.type.publication Research en_ZA
uct.type.resource Thesis en_ZA
dc.publisher.institution University of Cape Town
dc.publisher.faculty Faculty of Science en_ZA
dc.publisher.department Department of Molecular and Cell Biology en_ZA
dc.type.qualificationlevel Doctoral
dc.type.qualificationname PhD en_ZA
uct.type.filetype Text
uct.type.filetype Image
dc.identifier.apacitation Pfeffer, P. L. (1991). <i>The isolation and characterization of a P. Angulosus homeobox</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology. Retrieved from http://hdl.handle.net/11427/21983 en_ZA
dc.identifier.chicagocitation Pfeffer, Peter Lance. <i>"The isolation and characterization of a P. Angulosus homeobox."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1991. http://hdl.handle.net/11427/21983 en_ZA
dc.identifier.vancouvercitation Pfeffer PL. The isolation and characterization of a P. Angulosus homeobox. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Molecular and Cell Biology, 1991 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/21983 en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Pfeffer, Peter Lance AB - The aim of this thesis was to isolate and characterize a homeobox-containing gene of the South African sea urchin Parechinus angulosus. This was achieved by constructing a genomic library of several individuals and screening this library using a probe containing the Antennapedia homeobox. Eight clones were isolated and shown to represent different alleles of the same gene. One clone was sequenced, revealing a homeobox which was termed PaHboxl. This homeobox was compared to published homeobox sequences and shown to be a member of the Antp (Hoxl.l) subclass (table 1.1). A splice donor site was identified 23 bp upstream of the homeobox and the observation confirmed by RNAase mapping. PaHboxl is situated in a genomic area showing a significantly higher degree of restriction fragment polymorphism than expected. This was shown by a statistical analysis which should be of general value in the interpretation of such polymorphisms. The expression of PaHboxl was examined by RNAase protection assays and Northern blotting. Two distinct phases of expression were observed - during embryogenesis PaHboxl is expressed transiently at low levels in 11,5 hr mesenchyme blastula stage embryos (44 ± 8 transcripts per embryo) with levels 3-5 fold lower 2,5 hr before and after this stage. Expression is observed again at up to 160 fold higher levels in the adult with maximal expression in testis (11 transcripts per 10 pg total RNA), and increasingly lower levels in intestines, ovary and Aristotle's lantern. Two transcripts of size 5,2 and 5,7 kbp were observed. Expression in Aristotle's lantern and embryonic stages could not be detected by Northern analysis. DA - 1991 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1991 T1 - The isolation and characterization of a P. Angulosus homeobox TI - The isolation and characterization of a P. Angulosus homeobox UR - http://hdl.handle.net/11427/21983 ER - en_ZA


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