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Example sentences for: subcloned
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The adenovirus (AV) backbone for the aquaporin-1 (AQP1) sense and antisense AV constructs was a replication-deficient "first-generation" AV with deletions of the E1 and E3 genes [ 26 ] . This "empty" AV contains the cytomegalovirus (CMV) promoter and bovine growth hormone polyadenylation (bHG) site separated by a polylinker that was used to clone AQP1 DNA as described previously [ 27 ] . A recombinant AQP1 virus was constructed using a plasmid containing the coding sequence for AQP1, pCHIPev [ 28 ] . pCHIPev was digested and the AQP1 insert was subcloned into the shuttle vector pSKAC, creating pSKAC/AQP1.
The PCR-Select cDNA Subtraction and PCR-Select Differential Screening Kits (Clontech Laboratories Inc.) were used to enrich for and identify differentially expressed genes in the driver and tester cDNAs [ 48 49 ] . Both forward- and reverse-subtraction were performed; untreated HUVEC cDNA served as driver and VEGF-treated HUVEC cDNA served as tester in the former, whereas untreated HUVEC cDNA served as tester and VEGF-treated HUVEC cDNA served as driver in the latter [ 48 49 ] . Target cDNA fragments were amplified using the Marathon™ cDNA Amplification Kit, subcloned into pCR ®2.
Sequencing of subcloned genomic fragments and comparison with the cDNA sequence allowed us to construct an exon map of the NPM3 gene (Figure 1A).
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.
Research Genetics, Inc.) with RSAI and then subcloned the DNA fragments into pZero 2.0 according to the manufacturer's instructions.
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