Words similar to pet-gst-ppt

Example sentences for: pet-gst-ppt

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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.
PP5 was expressed and purified as described previously [ 18 ] . To generate the expression plasmid pET-GST-PPT1,
GAL1 promoter, a PPT1 DNA fragment was cut from pET-GST-PPT1 by EcoR I and Not I digestion and subcloned into the EcoR I/ Not I-digested YCpIF16 (ATCC No.
The cDNA probe was prepared by restriction digestion of pET-GST-PPT1 with Bsm I and Nco I. The 879 bp DNA fragment generated, corresponding to 544-1422 of PPT1 coding sequence, was gel-purified and labeled with 0.67 μM [α- 32P] CTP (3000 Ci/mmol) using the DECAprimeII random priming kit (Ambion) according to the user manual.