Sentences with phrase «rescuing unc»

An unc - 9:: gfp expression cassette was made by introduction of GFP (afmx) from the partially rescuing unc - 9:: gfp genomic construct (above).

(B) Fully rescuing unc - 7 min plasmid expressing UNC - 7S and UNC - 7SR.

A fully - rescuing unc - 7 construct (Figure 3B) extended to the Xho I site (nucleotide 15,574) and included a frameshift generated by filling in the Sal I site (nucleotide 14,412).

Our genomic unc - 9:: gfp construct was found to only partially rescue unc - 9 mutants.

Together these studies suggest that Punc - 7S drives expression of UNC - 7S in a subset of neurons (including AVA and AVB, but not motor neurons) that can effect rescue of forward locomotion, and promoter sequences upstream of exon 2 drive additional expression of UNC - 7S in motor neurons or possibly other neurons required to fully rescue all unc - 7 locomotory defects.

First, the genomic region represented by the unc - 9:: gfp construct was shown to fully rescue unc - 9 (fc16) animals, indicating that this region is sufficient to encode a functional unc - 9 product.

When expressed under control of the Punc - 7S promoter, UNC - 7L (Punc - 7S:: unc - 7L [M121L]-RRB- rescued unc - 7 (e5) forward locomotion; though much of the signal remained associated with cell bodies, some puncta were seen localized to AVB: B motor neuron gap junctions (Figure 10D).

M121 was mutated to L in unc - 7SΔ1S; this construct (unc - 7SΔ1S - M121L; Figure 3F) plus F56B12 failed to rescue unc - 7, suggesting that UNC - 7L does not have rescuing activity.

Not exact matches

(B) Pcex - 1:: unc - 7S:: gfp (AVA, AVD interneurons) rescues forward locomotion but does not concentrate in puncta near B cell bodies.

Lane 1, wild type (N2); lanes 2 and 3, unc - 7 (e5) rescued with unc - 7S:: gfp + cosmid F56B12, expected to express all isoforms; lane 4, unc - 7 (e5) rescued with unc - 7SΔ1S + F56B12, predicted to express UNC - 7L:: and SR:: GFP.

One rescued line, EH536, was maintained and used for genetic crosses involving unc - 9 (fc16).

Phenotypic rescue of unc - 7 by various constructs was assessed by tapping transformed animals on the nose or tail and observing whether or not smooth sinusoidal waves were propagated.

A shorter version of unc - 7 min lacked promoter sequences upstream of exon 2 (unc - 7S:: gfp; Figure 3C) and rescued forward but not backward locomotion, suggesting that promoter sequences in intron 3 (Punc - 7S; Figure 3C) drive a subset of UNC - 7S expression sufficient to rescue only forward movement.

We will refer to the predicted products of the unc - 7 rescuing transcript as UNC - 7S with the understanding that UNC - 7SR is also translated.

When co-injected with F56B12, the unc - 7S:: gfp construct rescued both forward and reverse locomotion.

Like unc - 7S:: gfp, this construct rescued forward locomotion in the absence of cosmid F56B12, and we postulated that M121 in exon IV (the canonical innexin start site) might be used to generate a shorter but functional UNC - 7S - like product (UNC - 7SR).

(D) UNC - 7L expressed in interneurons (Punc - 7S:: unc - 7L [M121L]-RRB- rescues forward locomotion in unc - 7 (e5) and localizes to puncta near B motor neuron cell bodies.

To demonstrate that mutation of M121 did not lead to loss of an essential protein function, exon 1S was added back to allow for expression of UNC - 7S (M121L), and this construct (unc - 7S - M121L; Figure 3G) fully rescued.

The unc - 9:: gfp construct partially rescued uncoordinated locomotion in unc - 9 (fc16)-- hermaphrodites exhibit wild - type forward movement interrupted with bouts of spastic kinking.

Using unc - 9 mutants and heterologous promoters, we showed that expression of UNC - 9 in B class motor neurons is necessary to rescue the localization of UNC - 7S expressed in AVB.

To address this, genetic mosaic analysis of rescue of forward locomotion by the unc - 7S construct was carried out.

Using anti-GFP antibodies, two major protein bands, not present in wild - type, were detected in unc - 7 (e5) animals rescued with unc - 7S:: GFP + F56B12 (Figure 3H).

The shortest genomic region capable of rescuing forward and backward locomotion defects in unc - 7 mutants encompassed the coding region of unc - 7 and 1 kb of promoter sequence upstream of exon 2 (unc - 7 min; Figure 3B).

Psra - 11:: unc - 7S:: gfp expressed in AVB did not rescue forward locomotion in e5 animals; the Pcex - 1:: unc - 7s: gfp construct, expressed in AVA and AVD but not AVB, did, however, rescue e5 forward locomotion.

Other strains used included CB1377 daf - 6 (e1377) X and CW129 unc - 9 (fc16) X. For transformation rescue, SP1531 ncl - 1 (e1865) unc - 36 (e251) III was sometimes used; the ncl - 1 gene was incidental in these cases.

In animals rescued with unc - 7SΔ1S + F56B12, predicted to express UNC - 7SR:: GFP but not UNC - 7S:: GFP, only the smaller band was detected.

Therefore, unc - 7 locomotion defects can be rescued by either UNC - 7S or UNC - 7SR, but UNC - 7L can not substitute for this function (although it may have a subtle role or function in another process).

It was surprising that re-establishment of AVB: B motor neuron gap junctions was not implicated in the rescue of unc - 7 (e5) forward locomotion by unc - 7S.

unc - 7 transformation rescue.

(C) Co-localization (arrow) in an unc - 7 (e5) animal rescued for forward locomotion with unc - 7S construct (Figure 3C, minus GFP).