Sentences with phrase «subcloned into»

Protein coding sequences were flanked with unique NotI and AscI restriction sites and subcloned into a derivative of the pTT3 expression plasmid between a 5» mouse variable κ light chain signal peptide [32], and a 3» tag consisting of the rat Cd4 domains 3 and 4 followed by an enzymatic biotinylation sequence and a hexahistidine tag [33].

The PCR product was cloned and the isolated Kpn I / Nhe I fragment then subcloned into a pBS sk - plasmid bearing a GFP (afmx) sequence with an Nhe I site at the translational start of GFP.

Selected Amrose variant fluorescent proteins were subcloned into the pCS2 + expression vector [32](EcoRI - XbaI) using the gel purified amplicons generated above.

All JAK1 and JAK2 mutants were generated using QuickChange XL II site - directed mutagenesis kit (Stratagene) and subcloned into the pMX - GFP (for JAK1) or pMEGIX - GFP (for JAK2) biscistronic retroviral vector upstream of the IRES as previously described.17 — 19 The mutated JAK1 and JAK2 constructs were verified by sequencing the full - length JAK1 and JAK2 using DYEnamic ET Dye Terminator Kit (Amersham Biosciences).

CCR5 cDNA was subcloned into the retroviral vector pBABE - puro.

pNIBSC1 was subcloned into pUC19 inframe with lacZ gene.

CXCR4 cDNA was subcloned into the retroviral vector pBABE - puro.

The PCR product was digested with NcoI and BamHI and subcloned into the pSLAX - 13 shuttle vector.

The amplified fragments were cloned into Topo vectors and later released with appropriate enzymes for subcloning into the NZTK2 vector.