Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system

The human Y -box protein I (YB- I) is a class of proteins involved in transcriptional and translational regulation of a wide range of genes. YB-1 is implicated in regulation of multiple cellular functions. Previous studies found that the YB-1 expression is upregulated in many tumours and is highl...

Full description

Bibliographic Details
Main Author: Wong Jian, Yuan
Format: Monograph
Language:English
Published: Universiti Sains Malaysia 2008
Subjects:
Online Access:http://eprints.usm.my/48911/
http://eprints.usm.my/48911/1/WONG%20JIAN%20YUAN%20-%2024%20pages.pdf
_version_ 1848881289934929920
author Wong Jian, Yuan
author_facet Wong Jian, Yuan
author_sort Wong Jian, Yuan
building USM Institutional Repository
collection Online Access
description The human Y -box protein I (YB- I) is a class of proteins involved in transcriptional and translational regulation of a wide range of genes. YB-1 is implicated in regulation of multiple cellular functions. Previous studies found that the YB-1 expression is upregulated in many tumours and is highly correlated with adverse cancer prognosis and resistance to cancer drugs. In order to further explore and understand the role of YB-1 in terms of transcriptional and translational regulation in development of malignancy tumours, an overexpression assay has been carried out. Two different recombinants YB- I plasmid, pGEX- 2TK-YB-1 and pREP-YB-1 were used to carry out over-expression assay in bacterial (Escherichia coli, BL-21) and mammalian system (He La cancer cells) respectively. The comparison was analyzed by western blot analysis using a YB-1 polyclonal antibody. The over-expression results show a different migration of YB-1 protein in bacterial and mammalian expression system compared to their actual theoretical size. Meanwhile, overexpression assay involved transfection of pREP-YB-1 into He La cells using calcium phosphate transfection method. Due to poor reproducibility and rare quantitative studies on the mechanisms of the calcium phosphate transfection, trouble shooting was done and the optimization steps were suggested in the discussion of this research.
first_indexed 2025-11-15T18:16:40Z
format Monograph
id usm-48911
institution Universiti Sains Malaysia
institution_category Local University
language English
last_indexed 2025-11-15T18:16:40Z
publishDate 2008
publisher Universiti Sains Malaysia
recordtype eprints
repository_type Digital Repository
spelling usm-489112021-04-18T02:54:21Z http://eprints.usm.my/48911/ Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system Wong Jian, Yuan R Medicine (General) The human Y -box protein I (YB- I) is a class of proteins involved in transcriptional and translational regulation of a wide range of genes. YB-1 is implicated in regulation of multiple cellular functions. Previous studies found that the YB-1 expression is upregulated in many tumours and is highly correlated with adverse cancer prognosis and resistance to cancer drugs. In order to further explore and understand the role of YB-1 in terms of transcriptional and translational regulation in development of malignancy tumours, an overexpression assay has been carried out. Two different recombinants YB- I plasmid, pGEX- 2TK-YB-1 and pREP-YB-1 were used to carry out over-expression assay in bacterial (Escherichia coli, BL-21) and mammalian system (He La cancer cells) respectively. The comparison was analyzed by western blot analysis using a YB-1 polyclonal antibody. The over-expression results show a different migration of YB-1 protein in bacterial and mammalian expression system compared to their actual theoretical size. Meanwhile, overexpression assay involved transfection of pREP-YB-1 into He La cells using calcium phosphate transfection method. Due to poor reproducibility and rare quantitative studies on the mechanisms of the calcium phosphate transfection, trouble shooting was done and the optimization steps were suggested in the discussion of this research. Universiti Sains Malaysia 2008 Monograph NonPeerReviewed application/pdf en http://eprints.usm.my/48911/1/WONG%20JIAN%20YUAN%20-%2024%20pages.pdf Wong Jian, Yuan (2008) Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system. Other. Universiti Sains Malaysia. (Submitted)
spellingShingle R Medicine (General)
Wong Jian, Yuan
Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title_full Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title_fullStr Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title_full_unstemmed Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title_short Over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
title_sort over-expression of human y-box binding proteini: a comparison between mammalian and bacterial expression system
topic R Medicine (General)
url http://eprints.usm.my/48911/
http://eprints.usm.my/48911/1/WONG%20JIAN%20YUAN%20-%2024%20pages.pdf