Whereas, in cluster-IIb PDB2
and PDB3 revealed 75% relatedness.
A mammalian expression vehicle (25) including pCMV-Int and pDB2, encoding PhiC31 integrase and EGFP coding region flanked by the cytomegalovirus promoter respectively, was kindly provided by Prof.
In order to construct a recombinant pDB2 plasmid expressing EGFP under the control of Oct4 promoter, a fragment encompassing the sequences of CMV promoter in pDB2 was removed by double digestion with NheI and VspI (Fermentas, Lithuania).
Finally, cloning of the amplicon was achieved by insertion into pDB2 vector in place of CMV promoter.
Both the new cell reporter and control cell [(stably transformed with pDB2 ([DELTA] promoter/EGFP) plasmid)] lines were treated by trypsin and assayed using a FACS with excitation at 488 nm for EGFP.
A 100 Amp Active Harmonic filter may be installed in PDB2 of Plant III supplied by TX1.The Plant III has got 30 Ring frames, each ring frame having VFD (Variable Frequency Drive) of 50 KW rating.
There were facing a Problem of High current harmonics more than 25% in PDB2.
The 100 Amp APF Unit may be installed across the load of PDB2.
Finally, total length fragment with VspI-NheI overhangs from pTZ57R/T was sub-cloned into pDB2 target vector (Figure 2).
To construct a promoter free vector, CMV promoter was pulled out from the backbone of pDB2 vector by a VspI-NheI double digestion.
When cells reached to 50-80% of confluency, transfection was carried out by pDB2/PPAR[gramma]1 promoter, pDB2 and promoter free pDB2 vectors by lipofectamine 2000 (Invitrogen) according to the manufacturer's instruction.