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Example sentences for: pet-

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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.
A fragment containing the PPT1 ORF was excised from the pBII-PPT1 using Eco RI and Xho I restriction enzymes and was subcloned downstream of the GST coding region into the Eco RI and Sal I sites of the bacterial expression vector pET-21a GST [ 37 ] . To construct the pET-GST-PPT1 (1-504) plasmid a 453 bp fragment was amplified from pET-GST-PPT1 by PCR using a 3' primer containing a Xho I restriction site and a stop codon designed to truncate the PPT1 gene product at Met504 (5'-gatcctcgagTTAcattggttttatatctggg) and a 5' primer (5'-gtaatgcatggtggttta) encompassing the Nsi I restriction site of PPT1 . The PCR product was cloned into the Nsi I and Xho I restriction sites of pET-GST-PPT1 and sequenced.
3, when induced with IPTG, the pET-15b-PfPP5 clone produced a polypeptide of approximate Mr 70 k (lane 2), which is in excellent agreement with the calculated MW of 69,139, considering that pET-15b adds a (His) 6 tag (about 2 kDa) to the N-terminus of the recombinant protein.
The expression vector pET-28a and E. coli expression strain BL21 (DE3) were obtained from Novagen.
To construct a pBS58 ftsZ mutant plasmid (pBS58 ftsZ mut), the EcoRI and BstEII fragment of ftsZ in pBS58 was replaced with the same part of ftsZ from pET- ftsZ mut.