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This finding means that an individual participant was more likely to call a foul when seeing a picture in its leftward compared to rightward orientation, even though the two stimuli were otherwise identical.
The activity levels induced by the piggyBac vectors containing the heterologous genes in the rightward orientation were significantly (5X to over 35X) higher.
Similarly, vectors encoding GalT, ST6GalI, and ST3GalIII genes in the rightward orientation induced about 8X, 8X, and 3X more activity, respectively, than vectors encoding these genes in the leftward orientation (Fig. 5, columns A-B).
For the observations on posture the 3D coordinates were defined as follows: the positive x direction was the forward orientation, the positive y direction was the rightward orientation, and the positive z direction was the upward orientation.
Thus, together with our previous results, these results suggested that the position effect observed with our new piggyBac vectors results from the activated expression of heterologous genes cloned in the rightward orientation, irrespective of the transcriptional control element.
Probe R containing the DNA sequence at nt 576 67 (complementary to RNAs transcribed in the rightward orientation) hybridized to the 350-nt RNA (ORF3) and the internal control band.
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A 1.2 kb StuI-SnaBI pH5-gfpzeo expression cassette from pG06dH5gz was blunt cloned in both leftward and rightward orientations into the Klenow-filled XbaI site in pMSΔ102 to yield plasmids pMSΔ102H5GZ.1, -.4, respectively.
Thus, this region was deleted from piggyBac vectors containing the constitutive GalT gene in the leftward or rightward orientations (Fig. 7, vectors B and D) and GalT expression levels induced by these new derivatives were compared to those induced by controls containing the intact promoter.
The enhanced green fluorescence protein (EGFP) coding sequence was cloned in both rightward and leftward orientations so as to replace TR-2.
Subsequently, transient expression assays were performed to compare the GalT expression levels induced by these new piggyBac derivatives with those induced by the original vectors encoding the constitutive or inducible GalT genes in either the rightward or leftward orientations.
This orientation would make a rightward step more favorable for the trailing head, resulting in a net rightward bias.
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