Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli

Two plasmids, pFLAG-ATS and pET 26b(+), were studied for the periplasmic expression of recombinant human interferon-α2b (IFN-α2b) in Escherichia coli. The pFLAG-ATS contains ompA signal sequence and tac promoter while pET 26b(+) contains pelB signal sequence and T7lac promoter. It was observed that...

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Main Authors: Ramanan, Ramakrishnan Nagasundara, Wong, Bee Tik, Memari, Hamid Rajabi, Azaman, Siti Nor Ani, Tau, Chuan Ling, Tey, Beng Ti, Mohd Lila, Mohd Azmi, Abdullah, Mohd Puad, Abdul Rahim, Raha, Ariff, Arbakariya
Format: Article
Language:English
Published: Academic Journals 2010
Online Access:http://psasir.upm.edu.my/id/eprint/11248/
http://psasir.upm.edu.my/id/eprint/11248/1/62.pdf
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author Ramanan, Ramakrishnan Nagasundara
Wong, Bee Tik
Memari, Hamid Rajabi
Azaman, Siti Nor Ani
Tau, Chuan Ling
Tey, Beng Ti
Mohd Lila, Mohd Azmi
Abdullah, Mohd Puad
Abdul Rahim, Raha
Ariff, Arbakariya
author_facet Ramanan, Ramakrishnan Nagasundara
Wong, Bee Tik
Memari, Hamid Rajabi
Azaman, Siti Nor Ani
Tau, Chuan Ling
Tey, Beng Ti
Mohd Lila, Mohd Azmi
Abdullah, Mohd Puad
Abdul Rahim, Raha
Ariff, Arbakariya
author_sort Ramanan, Ramakrishnan Nagasundara
building UPM Institutional Repository
collection Online Access
description Two plasmids, pFLAG-ATS and pET 26b(+), were studied for the periplasmic expression of recombinant human interferon-α2b (IFN-α2b) in Escherichia coli. The pFLAG-ATS contains ompA signal sequence and tac promoter while pET 26b(+) contains pelB signal sequence and T7lac promoter. It was observed that periplasmic expression of IFN-α2b from pET 26b(+) was around 3000 times higher than pFLAG-ATS. Difference in the expression level was attributed to the difference in the promoters and the signal sequences. In silico analysis of mRNA secondary structures were analyzed using Vienna RNA package and MFOLD. The resultssuggested that the increase of expression would mainly due to the difference in the translation initiation associated with secondary structure of mRNA transcribed by both plasmids.
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publishDate 2010
publisher Academic Journals
recordtype eprints
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spelling upm-112482017-11-14T02:48:15Z http://psasir.upm.edu.my/id/eprint/11248/ Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli Ramanan, Ramakrishnan Nagasundara Wong, Bee Tik Memari, Hamid Rajabi Azaman, Siti Nor Ani Tau, Chuan Ling Tey, Beng Ti Mohd Lila, Mohd Azmi Abdullah, Mohd Puad Abdul Rahim, Raha Ariff, Arbakariya Two plasmids, pFLAG-ATS and pET 26b(+), were studied for the periplasmic expression of recombinant human interferon-α2b (IFN-α2b) in Escherichia coli. The pFLAG-ATS contains ompA signal sequence and tac promoter while pET 26b(+) contains pelB signal sequence and T7lac promoter. It was observed that periplasmic expression of IFN-α2b from pET 26b(+) was around 3000 times higher than pFLAG-ATS. Difference in the expression level was attributed to the difference in the promoters and the signal sequences. In silico analysis of mRNA secondary structures were analyzed using Vienna RNA package and MFOLD. The resultssuggested that the increase of expression would mainly due to the difference in the translation initiation associated with secondary structure of mRNA transcribed by both plasmids. Academic Journals 2010-01-18 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/11248/1/62.pdf Ramanan, Ramakrishnan Nagasundara and Wong, Bee Tik and Memari, Hamid Rajabi and Azaman, Siti Nor Ani and Tau, Chuan Ling and Tey, Beng Ti and Mohd Lila, Mohd Azmi and Abdullah, Mohd Puad and Abdul Rahim, Raha and Ariff, Arbakariya (2010) Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli. African Journal of Biotechnology, 9 (3). art. no. E05979530638. pp. 285-292. ISSN 1684–5315 http://www.academicjournals.org/journal/AJB/article-abstract/E05979530638
spellingShingle Ramanan, Ramakrishnan Nagasundara
Wong, Bee Tik
Memari, Hamid Rajabi
Azaman, Siti Nor Ani
Tau, Chuan Ling
Tey, Beng Ti
Mohd Lila, Mohd Azmi
Abdullah, Mohd Puad
Abdul Rahim, Raha
Ariff, Arbakariya
Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title_full Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title_fullStr Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title_full_unstemmed Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title_short Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli
title_sort effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in escherichia coli
url http://psasir.upm.edu.my/id/eprint/11248/
http://psasir.upm.edu.my/id/eprint/11248/
http://psasir.upm.edu.my/id/eprint/11248/1/62.pdf