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OJVRTM
Online Journal of Veterinary Research©
Volume 25 (3): 207-213,
2021.
Lentiviral gene vector transformation in Escherichia
coli derived H5α, JM109 and Stbl4 cells
Azam Mokhtari1, 2, Shekoofeh Rezayi1
1Department of
Pathobiology, Faculty of Veterinary Medicine, 2Research Institute of
Zoonotic Diseases, Shahrekord University, Shahrekord, Iran
ABSTRACT
Mokhtari A, Rezayi S., Lentiviral gene vector transformation in Escherichia coliderived
H5α, JM109 and Stbl4 cells, Onl J Vet Res., 25 (3):207-213, 2021.
Bacterial cell transformation is essential for gene cloning. Although
plasmid gene transfer recombinant lentivectors are
highly effective, they do not transform readily in most cells. E coli
derived Stbl2 and Stbl3 strains maintain
structure of lentiviral plasmid pRRL.SIN.cPPT.PGK/Oligo2-IRES-DsRed-WPRE
and boost colony growth. However, Stbl4 appears to be even more efficient to
justify further study. We thus investigated plasmid transformation in DH5α,
JM109 and Stbl4 cells derived from Escherichia coli. To transform
recombinant plasmids in DH5α, Stbl4 and JM109 and confirm the process we
used negative controls without plasmid and a positive control by heat shock
method with Amp resistance gene. We calculated
transformation cloning units by bacterial suspension culture under
identical cell conditions and ran 20 replications for each cell. Calcium
chloride precipitation and thermal shock was done to determine transform
efficiency by colonies yielding 0.26 × 106 for DH5α,
0.89 × 106 for JM109 and 1.57 × 106 for Stbl4 cfu / µg. Our
results suggest that optimal cell for pWPI lentiviral
vector was DH5α and Stbl4 most suitable for recombinant pCDH vectors.
Keywords: Transformation,
competent cell, DH5α, JM109, Stbl4, colony formation unit.
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