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Paper
Title

Establishment a CHO Cell Line Expressing Human CD۵۲ Molecule

Year: 1395
COI: JR_RBMB-5-1_009
Language: EnglishView: 49
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Authors

Khadijeh Tati - Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran - Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Mahsa Yazdanpanah-Samani - Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran
Amin Ramezani - Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran - Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran
Elham Mahmoudi Maymand - Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran
Abbas Ghaderi - Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran

Abstract:

Background: CD۵۲ is a small glycoprotein with a GPI anchor at its C-terminus. CD۵۲ is expressed by Normal and malignant T and B lymphocytes and monocytes. There are detectable amounts of soluble CD۵۲ in plasma of patients with CLL and could be used as a tumor marker. Although the biological function of CD۵۲ is unknown but it seems that CD۵۲ may be involved in migration and activation of T-cells .The aim of this study was to clone and express human CD۵۲ gene in CHO cell line and studying its function in more details. Methods: Based on GenBank databases two specific primers were designed for amplification of cd۵۲ gene. Total RNA was extracted from Raji cell line and cDNA synthesized. Amplified fragment was cloned in pBudCE۴.۱ vector. The new construct was transfected to CHO-K۱ cell line using electroporation method. Expression of recombinant CD۵۲ protein was evaluated by Real time PCR and flow cytometry methods. Results: Amplification of CD۵۲ gene using specific primers on Raji cDNA showed a ۲۰۹ bp band. New construct was confirmed by PCR and restriction pattern and sequence analysis. The new construct was designated as pBudKT۱. RT-PCR analysis detected cd۵۲ mRNAs in transfected cells and Flow cytometry Results showed that ۷۸.۴ % of cells represented CD۵۲ in their surfaces. Conclusions: In conclusion, we established a human CD۵۲ positive cell line, CHO-CD۵۲, and the protein was expressed on the membrane. Cloning of the CD۵۲ gene could be the first step for the production of therapeutic monoclonal antibodies and detection systems for soluble CD۵۲ in biological fluids.

Keywords:

CD۵۲ , Recombinant DNA , Therapeutic and diagnostic proteins

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This Paper COI Code is JR_RBMB-5-1_009. Also You can use the following address to link to this article. This link is permanent and is used as an article registration confirmation in the Civilica reference:

https://civilica.com/doc/1263073/

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Tati, Khadijeh and Yazdanpanah-Samani, Mahsa and Ramezani, Amin and Maymand, Elham Mahmoudi and Ghaderi, Abbas,1395,Establishment a CHO Cell Line Expressing Human CD۵۲ Molecule,https://civilica.com/doc/1263073

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Type of center: علوم پزشکی
Paper count: 2,546
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