Medical Journal of The Islamic Republic of Iran (MJIRI)
Iran University of Medical Sciences
Thu, Dec 12, 2024
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Volume 12, Issue 2 (8-1998)
Med J Islam Repub Iran 1998 |
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SHAH-HOSSEINY M, AKBARI M, TABARRAI B, RECHINSKY V. PCR-MEDIATED CLONING A ND EXPRESSION OF THE GENE FOR THE B-SUBUNIT OF VIBRIO CHOLERAE TOXIN ISOLATED RECENTLY IN IRAN. Med J Islam Repub Iran 1998; 12 (2) :123-128
URL: http://mjiri.iums.ac.ir/article-1-1017-en.html
URL: http://mjiri.iums.ac.ir/article-1-1017-en.html
From the Biology Unit, Faculty of Science, Imam Hossein University, Tehran
Abstract: (4148 Views)
Knowing the nucleotide sequence of the cholera toxin operon, we designed
oligonucleotide primers for its-PCR amplification from local clinical isolates of
V. cholerae. The resulting amplification product was cloned in a common
pUC18 vector. Subsequently, a part of this operon encoding the cholera toxin Bsubunit
(CTB) was reamplified and cloned between the BamH1 and EcoR1 sites
of the same vector to create a recombinant plasmid pR18CTB. Temperaturecontrolled
expression of the target protein was achieved by supplementing
pR18CTB with a DNA fragment which contained a strong promoter PR and the
gene for a heat-sensitive repressor cI857 of bacteriophage lambda from a n
expression vector pCQV2. When induced, the constructed plasmid pSCTB18
provided for the production of recombinant CTB secreted into the periplasmic
space in a yield of about 3mg per liter of bacterial culture, as revealed by OM 1-
ELISA.
Type of Study: Original Research: Basic Science in Medicine |
Subject:
Biological Sciences
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