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Reports of Biochemistry and Molecular Biology
rbmb.net
Basic Sciences
http://rbmb.net
1
admin
2322-3480
2322-3480
10.61882/rbmb
en
jalali
1395
7
1
gregorian
2016
10
1
5
1
online
1
fulltext
en
Establishment a CHO Cell Line Expressing Human CD52 Molecule
زیست شناسی سلولی
Cell Biology
<p style="text-align: justify;"><em><strong>Background: </strong></em>CD52 is a small glycoprotein with a GPI anchor at its C-terminus. CD52 is expressed by Normal and malignant T and B lymphocytes and monocytes. There are detectable amounts of soluble CD52 in plasma of patients with CLL and could be used as a tumor marker. Although the biological function of CD52 is unknown but it seems that CD52 may be involved in migration and activation of T-cells .The aim of this study was to clone and express human CD52 gene in CHO cell line and studying its function in more details.</p>
<p style="text-align: justify;"><em><strong>Methods:</strong></em> Based on GenBank databases two specific primers were designed for amplification of <em>cd52</em> gene. Total RNA was extracted from Raji cell line and cDNA synthesized. Amplified fragment was cloned in pBudCE4.1 vector. The new construct was transfected to CHO-K1 cell line using electroporation method. Expression of recombinant CD52 protein was evaluated by Real time PCR and flow cytometry methods.</p>
<p style="text-align: justify;"><strong><em>Results:</em> </strong>Amplification of CD52 gene using specific primers on Raji cDNA showed a 209 bp band. New construct was confirmed by PCR and restriction pattern and sequence analysis. The new construct was designated as pBudKT1. RT-PCR analysis detected cd52 mRNAs in transfected cells and Flow cytometry Results showed that 78.4 % of cells represented <em>CD52</em> in their surfaces.</p>
<p style="text-align: justify;"><em><strong>Conclusions: </strong></em>In conclusion, we established a human CD52 positive cell line, CHO-CD52, and the protein was expressed on the membrane. Cloning of the CD52 gene could be the first step for the production of therapeutic monoclonal antibodies and detection systems for soluble CD52 in biological fluids.</p>
CD52, Recombinant DNA, Therapeutic and diagnostic proteins
56
61
http://rbmb.net/browse.php?a_code=A-10-1-66&slc_lang=en&sid=1
Khadijeh
Tati
10031947532846006188
10031947532846006188
No
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
10031947532846006189
10031947532846006189
No
Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran
Amin
Ramezani
10031947532846006190
10031947532846006190
No
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
10031947532846006191
10031947532846006191
No
Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran
Abbas
Ghaderi
ghaderia@sums.ac.ir
10031947532846006192
10031947532846006192
Yes
Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran