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Example sentences for: pet-

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The (His) 6 -tagged PfPP1 expressed from pET-15b-PfPP1 was purified through Ni +2-chelation chromatography [ 14 ] as described by the manufacturer (Novagen), with 1 mM MnCl 2 being present in all the buffers.
A His-tag at the N-terminal of the p66+6aa subunit was introduced by sub cloning the Bal-I and Hind III fragment of pKKRT66+6aa into pET-28a-RT66 expression cassette.
The PCR products were gel-purified using low melting temperature agarose and subjected to restriction enzyme digestion with Nde I and Xho I. The same two restriction enzymes were used to digest plasmid pET-24b+ DNA.
3, a protein band of approximately 37,000 Mr was produced from the pET-15b-PfPP1 clone, which is in agreement with the predicted molecular weight of 34,904 of PfPP1 and roughly another 2,000 added for the (His) 6 region of pET-15b (Novagen: Madison, WI).
Growth and induction of E. coli BL21(DE3) containing pET-15b-PfPP1 and the RIG plasmid were carried out using procedures described earlier [ 14 48 ] , except that the culture was grown at 18°C in the presence of 2 mM MnCl 2 , and IPTG concentration was lowered to 0.4 mM.