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UBC Theses and Dissertations
Sequencing adjacent to BssHII sites on human chromosome 8 : a method of gene identification Debella, Leah Rae
Abstract
Considerable research is focused on the identification of genes in the human genome. Recently, the Human Genome project has named gene identification as one of its goals to be accomplished in tandem with mapping and sequencing of the entire genome. As a result,a large program centered on the creation of expressed sequence tags (ESTs) is currently underway. Although this method of gene identification has vastly increased the number of expressed sequences in the database, it is unlikely that all genes encoded in the genome with be detected though the sequencing of cDNAs. An alternative method of gene identification, using genomic DNA, is the sequencing of regions adjacent to rare cutting enzyme sites. Recognition sites of rare cutting enzymes are found infrequently in genomic DNA but cluster in regions known as CpG islands. CpG islands are associated with most ubiquitously expressed genes and 40% of tissue specific genes. Therefore, sequencing of these regions offers a complementary method to be used in the recognition of novel genes. The objective of this project was to identify novel genes present in cloned DNA from human chromosome 8 using BssHII, a rare cutting enzyme.BssHII, is an excellent choice to be used in this manner, as 80% of its sites are located in CpG islands. DNA adjacent to fourteen Bssrlll sites, originating from eleven cosmids, was sequenced. From this set two human ESTs were identified; two open reading frames with no apparent homologues; two CpG islands, one of which contains a translation initiation signal and is 5' of the EST H88121; and two novel human genes. These results validate the hypothesis that the use of this method can complement other techniques in the identification of novel genes in the human genome.
Item Metadata
Title |
Sequencing adjacent to BssHII sites on human chromosome 8 : a method of gene identification
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
1997
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Description |
Considerable research is focused on the identification of genes in the human
genome. Recently, the Human Genome project has named gene identification as one of its
goals to be accomplished in tandem with mapping and sequencing of the entire genome.
As a result,a large program centered on the creation of expressed sequence tags (ESTs) is
currently underway. Although this method of gene identification has vastly increased the
number of expressed sequences in the database, it is unlikely that all genes encoded in the
genome with be detected though the sequencing of cDNAs.
An alternative method of gene identification, using genomic DNA, is the
sequencing of regions adjacent to rare cutting enzyme sites. Recognition sites of rare
cutting enzymes are found infrequently in genomic DNA but cluster in regions known as
CpG islands. CpG islands are associated with most ubiquitously expressed genes and
40% of tissue specific genes. Therefore, sequencing of these regions offers a
complementary method to be used in the recognition of novel genes.
The objective of this project was to identify novel genes present in cloned DNA
from human chromosome 8 using BssHII, a rare cutting enzyme.BssHII, is an excellent
choice to be used in this manner, as 80% of its sites are located in CpG islands. DNA
adjacent to fourteen Bssrlll sites, originating from eleven cosmids, was sequenced. From
this set two human ESTs were identified; two open reading frames with no apparent
homologues; two CpG islands, one of which contains a translation initiation signal and is
5' of the EST H88121; and two novel human genes. These results validate the hypothesis
that the use of this method can complement other techniques in the identification of novel
genes in the human genome.
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Extent |
9175476 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-03-24
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0087937
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
1997-11
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.