Volume 12, Issue 2 (8-1998)
Med J Islam Repub Iran 1998 |
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From the Biology Unit, Faculty of Science, Imam Hossein University, Tehran
Abstract: (4367 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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