Cloning and expression of truncated CTCF
The CTCF gene maps on chromosome 16 at 16q21- q22.3. It encodes a transcriptional factor protein CTCF called zinc finger protein. CTCF has a number of functions in the cell including controls of cell proliferation by having interactions with other proteins. There are several proteins associated w...
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| Format: | Article |
| Language: | English |
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Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia
2005
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| Online Access: | http://eprints.usm.my/47761/ http://eprints.usm.my/47761/1/CHOT%20SAN%20NGAN-24%20pages.pdf |
| _version_ | 1848880966906413056 |
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| author | Ngan, Chot San |
| author_facet | Ngan, Chot San |
| author_sort | Ngan, Chot San |
| building | USM Institutional Repository |
| collection | Online Access |
| description | The CTCF gene maps on chromosome 16 at 16q21- q22.3. It encodes a transcriptional
factor protein CTCF called zinc finger protein. CTCF has a number of functions in the
cell including controls of cell proliferation by having interactions with other proteins.
There are several proteins associated with CTCF. Among them are proteins involved in
transcriptional and cell proliferation control, RNA processing, signal transduction,
nucleosome components and tumor suppressors.
CTCF consists of N-tenninal domain, zinc finger domain and C-terminal domain.
Extensive research have been done to elucidate the important regions that might play
important role in both DNA binding and protein-protein interactions. Previous studies has
shown there was a region in the C-domain which has direct interaction with large subunit
of RNA polymerase IT and controlling various cellular process.
In this study, a truncated CTCF from the C-tenninal domain was produced by polymerase
chain reaction (PCR). The amplified products was then subcloned into the intermediate
cloning vector, pTOP02.1 and transformed into E. coli strain DH5a. The insert was then
cut with respective restriction enzyme to obtain the truncated region and further ligated
into pET16b expression vector for its expression in E. coli strain BL21 (DE3). Expressed
protein was separated using SDS-PAGE followed by Western Blotting using a-Histag
monoclonal antibody. The truncated CTCF protein was detected to migrate at the size of
approximately 27kDa despite its theoretical size of 4kDa. This migration was expected
and due to anomalous conformational changes during migration. |
| first_indexed | 2025-11-15T18:11:32Z |
| format | Article |
| id | usm-47761 |
| institution | Universiti Sains Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T18:11:32Z |
| publishDate | 2005 |
| publisher | Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | usm-477612020-10-26T04:09:56Z http://eprints.usm.my/47761/ Cloning and expression of truncated CTCF Ngan, Chot San R Medicine The CTCF gene maps on chromosome 16 at 16q21- q22.3. It encodes a transcriptional factor protein CTCF called zinc finger protein. CTCF has a number of functions in the cell including controls of cell proliferation by having interactions with other proteins. There are several proteins associated with CTCF. Among them are proteins involved in transcriptional and cell proliferation control, RNA processing, signal transduction, nucleosome components and tumor suppressors. CTCF consists of N-tenninal domain, zinc finger domain and C-terminal domain. Extensive research have been done to elucidate the important regions that might play important role in both DNA binding and protein-protein interactions. Previous studies has shown there was a region in the C-domain which has direct interaction with large subunit of RNA polymerase IT and controlling various cellular process. In this study, a truncated CTCF from the C-tenninal domain was produced by polymerase chain reaction (PCR). The amplified products was then subcloned into the intermediate cloning vector, pTOP02.1 and transformed into E. coli strain DH5a. The insert was then cut with respective restriction enzyme to obtain the truncated region and further ligated into pET16b expression vector for its expression in E. coli strain BL21 (DE3). Expressed protein was separated using SDS-PAGE followed by Western Blotting using a-Histag monoclonal antibody. The truncated CTCF protein was detected to migrate at the size of approximately 27kDa despite its theoretical size of 4kDa. This migration was expected and due to anomalous conformational changes during migration. Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia 2005-03 Article NonPeerReviewed application/pdf en http://eprints.usm.my/47761/1/CHOT%20SAN%20NGAN-24%20pages.pdf Ngan, Chot San (2005) Cloning and expression of truncated CTCF. Cloning and expression of truncated CTCF. (Submitted) |
| spellingShingle | R Medicine Ngan, Chot San Cloning and expression of truncated CTCF |
| title | Cloning and expression of truncated
CTCF |
| title_full | Cloning and expression of truncated
CTCF |
| title_fullStr | Cloning and expression of truncated
CTCF |
| title_full_unstemmed | Cloning and expression of truncated
CTCF |
| title_short | Cloning and expression of truncated
CTCF |
| title_sort | cloning and expression of truncated
ctcf |
| topic | R Medicine |
| url | http://eprints.usm.my/47761/ http://eprints.usm.my/47761/1/CHOT%20SAN%20NGAN-24%20pages.pdf |