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17942

Construction, transfection and production of recombinant vigilin in mammalian expression system

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Last updated: 03 Jan 2025

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Abstract

Vigilin is an abundant, highly conserved, ubiquitous protein containing 15 related, but non-identical, K-homolous nucleic acid binding domains. The construction, transfection and production of recombinant vigilin in mammalian expression system were investigated. The whole length of vigilin was amplified by polymerase chain reaction (PCR) and ligated to pCEP- PU vector. The recombinant construct pCEP-PU with vigilin was produced and transfected into Human embryonic kidney cells in a specific culture medium. The secreted recombinant vigilin was purified from the serum-free medium to homogeneity by affinity chromatography. The conditioned and purified media were tested for the presence of vigilin by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting with specific antibody. An immunoreactive band with an apparent molecular mass of approximately 140 kDa was detected. Immunofluorescence staining of transfected cells with vigilin demonstrated that recombinant vigilin molecules are localized in the nucleus and cytoplasm. In summary, the purified recombinant vigilin will facilitate future studies that address the structure and function of vigilin.

DOI

10.21608/ejhm.2006.17942

Keywords

SDS-PAGE/vigilin/Immunoblott/Immunofluorescence/affinitychromatography

Authors

First Name

Sayed Kamel

Last Name

Areida

MiddleName

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Affiliation

Zoology Department, Faculty of Science, Mansoura University, Mansoura, Egypt.

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Volume

23

Article Issue

1

Related Issue

3738

Issue Date

2006-04-01

Receive Date

2018-10-30

Publish Date

2006-04-01

Page Start

309

Page End

318

Print ISSN

1687-2002

Online ISSN

2090-7125

Article File

EJHM_Volume 23_Issue 1_Pages 309-318.pdf

PDF . 607.6kB

Link

https://ejhm.journals.ekb.eg/article_17942.html

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https://ejhm.journals.ekb.eg/service?article_code=17942

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12

Type

Original Article

Type Code

606

Publication Type

Journal

Publication Title

The Egyptian Journal of Hospital Medicine

Publication Link

https://ejhm.journals.ekb.eg/

MainTitle

Construction, transfection and production of recombinant vigilin in mammalian expression system

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Article

Created At

22 Jan 2023