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Cloning and expression of muc-1 gene in prokaryotic host for early detection of breast cancer
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Marjaneh Kazemi1   , Jafar Amani2 , Ali Hatef Salmanian3 , Elahe Gheybi4 |
1- Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
2- Associate Professor, Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. , jafar.amani@gmail.com
3- Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
4- Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran. |
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Abstract: (7405 Views) |
| Background and Aim: Breast cancer is the second most common cause of death among women in the world. Normal mammary gland and breast carcinomas are under the control of regulatory factors, activators and inhibitors inside the breast tissue, as well as a number of growth factors, receptors and proteins outside the breast tissue. Levels of tumor-associated antigens can be used as a predictor in the treatment of this disease. Use of antibodies against MUC1 antigen which is over expressed in 90% of breast cancers is a modern method of treatment. MUC1 tumor antigen disturbs the function of E-cadherin as a cell adhesion molecule. The purpose of this study was to produce MUC-1 recombinant protein for early diagnosis of breast cancer. Material and Methods: A part of MUC-1 gene was amplified by PCR. Then it was cloned into plasmid pET28a in order to be expressed in prokaryotic system. Plasmid pET28a was entered into E.coli BL21DE3 using heat shock method. Cloning process by digestive enzymes and sequence determination were confirmed. Bacteria containing recombinant plasmids were induced by using IPTG and the protein expression was investigated by SDS-PAGE gel. Results: The gene was cloned in the plasmid and the method was confirmed by enzyme digestion and sequencing. Gene expression was confirmed by western blotting. Conclusion: A part of human recombinant MUC-1 gene was produced in E.coli bacteria which can be used as a suitable diagnostic candidate for breast cancer. Keywords: Cloning, MUC-1, Expression Received: May 7, 2014 Accepted: Jun 29, 2015 |
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| Keywords: Cloning, MUC-1, Expression |
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Full-Text [PDF 1359 kb]
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Type of Study: Original Research |
Subject:
General
Received: 2015/10/6 | Accepted: 2015/10/6 | Published: 2015/10/6
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