Medical Journal of the Islamic Republic Of Iran
مجله پزشکی جمهوری اسلامی ایران
Med J Islam Repub Iran
Medical Sciences
http://mjiri.iums.ac.ir
2
journal2
1016-1430
2251-6840
8
10.18869/mjiri
14
8888
13
en
jalali
1377
5
1
gregorian
1998
8
1
12
2
online
1
fulltext
en
PCR-MEDIATED CLONING A ND EXPRESSION OF THE GENE FOR THE B-SUBUNIT OF VIBRIO CHOLERAE TOXIN ISOLATED RECENTLY IN IRAN
Biological Sciences
Biological Sciences
Original Research: Basic Science in Medicine
Original Research: Basic Science in Medicine
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.
V. cholerae, toxin, cloning, PCR
123
128
http://mjiri.iums.ac.ir/browse.php?a_code=A-10-298-468&slc_lang=en&sid=1
MH
SHAH-HOSSEINY
20031947532846006855
20031947532846006855
Yes
From the Biology Unit, Faculty of Science, Imam Hossein University, Tehran
MR
AKBARI
20031947532846006856
20031947532846006856
No
the Biochemistry Unit, Pasteur Institute of Iran
B
TABARRAI
20031947532846006857
20031947532846006857
No
the Biochemistry Unit, Pasteur Institute of Iran
V
RECHINSKY
20031947532846006858
20031947532846006858
No
the Office for Scientific and Industrial Studies, Presidency, Tehran, I.R. Iran.