Isolation and characterisation of the 5' region of the bovine lactoferrin gene : a dissertation presented to Massey University in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistry

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dc.contributor.authorBain, Heather B
dc.date.accessioned2012-03-13T01:25:01Z
dc.date.available2012-03-13T01:25:01Z
dc.date.issued1995
dc.description.abstractLactoferrin is an iron-binding protein found in a variety of mammalian secretions and polymorphonuclear leucocytes. Many biological roles have been proposed for lactoferrin however despite extensive research, the precise function(s) of this protein remain unclear. Lactoferrin appears to be regulated in a highly specific manner, dependent upon the developmental stage, the tissue and the species being investigated. RNA analyses have suggested that transcriptional regulation may be a significant factor controlling the expression of bovine lactoferrin within the mammary gland. A detailed knowledge of the molecular mechanisms and factors involved in the transcriptional regulation of the bovine lactoferrin gene will provide insights into the biological role of this protein. It was with this primary aim in mind that an unamplified bovine genomic library was screened for the presence of bovine lactoferrin 5' regulatory sequences. This resulted in the isolation of a clone which was characterised by detailed restriction mapping and found to contain sequences complementarity to bovine lactoferrin exon I and exon II sequences. Analysis of this clone indicated that is contained ~ 10 kb of DNA 5' to the transcriptional start site of bovine lactoferrin. A ~2.85 kb fragment which hybridised to an exon I probe was isolated and subjected to dideoxy sequencing. ~2.5 kb of DNA 5' to the lactoferrin coding sequence was identified within this fragment. A computer sequence homology search indicated that several putative consensus binding sites for transcription factors may be located within this DNA sequence. Fragments prepared from this subclone by PCR were subcloned into reporter gene expression constructs. The transcriptional activity of one of these constructs was investigated using COS cells and transient reporter gene expression studies. This construct, containing 565 bp of 5' bovine lactoferrin sequence, exhibited promoter activity. The successful isolation of this promoter will enable further investigations directed towards elucidating the transcriptional regulation of the bovine lactoferrin gene to be performed.en_US
dc.identifier.urihttp://hdl.handle.net/10179/3152
dc.language.isoenen_US
dc.publisherMassey Universityen_US
dc.rightsThe Authoren_US
dc.subjectLactoferrinen_US
dc.subjectGenetic aspectsen_US
dc.titleIsolation and characterisation of the 5' region of the bovine lactoferrin gene : a dissertation presented to Massey University in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Biochemistryen_US
dc.typeThesisen_US
massey.contributor.authorBain, Heather B
thesis.degree.disciplineBiochemistry
thesis.degree.grantorMassey University
thesis.degree.levelDoctoral
thesis.degree.levelDoctoralen
thesis.degree.nameDoctor of Philosophy (Ph.D.)
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