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Research in Pharmaceutical Sciences، جلد ۱۴، شماره ۴، صفحات ۳۵۱-۳۵۸
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی Evidence for expression of promoterless GFP cassette: Is GFP an ideal reporter gene in biotechnology science?
چکیده انگلیسی مقاله Green fluorescent protein (GFP) has played an important role in biochemistry and cell biology as a reporter gene. It has been used to assess the potency of promoters for recombinant protein production.This investigation reveals evidences suggesting that the gfp GFP gene (EGFP) could be expressed without the promoter. In a study, a pLenti-F/GFP vector was constructed with the purpose to allow GFP expression in transduced cells but not in packaging cells; however, after transfecting the HEK293T cell line, GFP gene was expressed, compared to pLOX/CWgfp-transfected cells showed expression lag, lower levels and reduced percentage of GFP expression in the cells. This unexpected result could be due to auto transduction in packaging cell, possible retrotransposon activity in the cell line, possible contamination of pLenti-F/GFP with the pLOX/CWgfp and possible presence of a promoter within backbone of the vector. All the possibilities were ruled out. To exclude the possibility that a sequence within the region might act as a promoter, the fragment to be transfected was minimized to a region containing “from the start of the GFP gene to 5’LTR R”. The GFP gene was again expressed. Therefore, our findings suggest the EGFP does not need promoter for expression. This should appeal to the researchers designing GFP based assays to evaluate the potency of promoters, since possible aberrant expression may have a potential to influenceon the results of a planned experiment.
کلیدواژه‌های انگلیسی مقاله Green fluorescent proteins, Viral packaging, Promoterless.
نویسندگان مقاله | Zahra Mohammadi
1Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of M


| Arezou Karamzadeh
1Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran. 3Pediatric Inherited Diseases Research Center, Research Institute for Primordial Prevention of Non-communicable Disease, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Mohammad Amin Tabatabaiefar
1Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran. 2Applied Physiology Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Hossein Khanahmad
4Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, I.R. Iran. 5Department of Biomaterials, Nanotechnology and Tissue Engineering, School of Advanced Technologies in Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.


| Laleh Shariati


نشانی اینترنتی http://rps.mui.ac.ir/index.php/jrps/article/view/1916
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/115/article-115-1870959.pdf
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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