Several of the sequences were very short products (three sequences), and nine of the remaining 12 insert sequences were oriented in the forward orientation relative to the Lac promoter of the bacterial expression plasmid.
The reverse orientation produced superior granule size distribution with improved liquid distribution attributes compared to the forward orientation.
The nature of the orientation has been determined by HPLC that reveals that the new analog incorporates exclusively in the forward orientation.
The recombinant clones having the specific insert in the forward orientation was then sequenced from both directions to get a contiguous sequence in an automated sequencer (ABI prism, model 377, Version 3.0).
With double-stranded DNA, there are six possible reading frames, three in the forward orientation on one strand and three reverse on the opposite strand.
Clones with the insert in the forward orientation were identified by nucleotide sequence analysis.
A clone (A1) containing the ADAMTS-1 cDNA in the forward orientation was subsequently identified by DNA sequencing.
Those with the 3' Bam segment in the forward orientation generally expressed cell surface CD2 protein at higher levels on PBL, despite their lower copy number (Figure 2A).
Northern blot analysis showed that mRNAs from cells transfected with the renilla/firefly dfoxO 5'UTR-B inserted in forward orientation were similar in size to those with the dfoxO 5'UTR-B in reverse orientation (Fig. 3C).
The predicted ORFs were not distributed equally on both strands; 71 were in forward orientation whereas 48 were in the reverse orientation, with many of these being arranged in unidirectional gene clusters.
