For the MS sequence data, the combined set of polymorphisms observed for "forward" and "reverse" orientation reads was filtered on G to A and C to T substitutions only, as these are the changes expected from EMS-induced treatment.
Both seem to be active, as the forward strand reads prevail immediately downstream of the first, forward promoter, while reverse orientation reads originate from the second, reverse strand promoter.
A good orientation involves reading a little history – you know, history, the part of the guidebook you usually skip past – and soaking up two powerful museums that go a long way to explaining Paraguayan identity.
Restriction digests and DNA sequencing confirmed the correct orientation and reading frames of the constructs.
Meanwhile, because the production of the constructs is laborious and often hampered by inappropriately positioned restriction sites, we also adopted Gateway cloning technology to join fragments in a predefined order, orientation, and reading frame [19].
These systems allow the correct orientation and reading frame to be maintained after transfer.
The new recombinants were then sequenced to verify the identity, proper orientation, and reading frame by J. C. Martinez-Cruzado (University of Puerto Rico, Mayagüez, PR).
Group 1 (self-learning reading, n=120) participants received orientation on reading materials to learn knowledge on hypertension through the poster text messages on blackboards and health education booklets monthly.
Five hundred and twenty six (85%) of the lines carried insertions into introns between protein coding exons in the orientation and reading frame expected for bona fide protein traps, generating protein traps in 346 unique genes.
The sequence, orientation, and reading frame were further determined and confirmed by sequencing analysis shown in Figure 3. Expression of target protein was checked by Coomassie blue staining of SDS-PAGE and immunoblotting.
Only a small percentage of the transpositions function as protein traps, because the insertion must be within the intron between two protein-coding exons, as well as being in the right orientation and reading frame to create an in-frame protein fusion when the artificial exon is spliced into the mRNA of the inserted gene.
