WormMine: ExpressionPattern Expr3459

• Pattern: Both transgenes were expressed in pharyngeal neurons and muscle, ventral nerve cord (VNC), vulval neurons and muscle, tail neurons, and spermatheca sheath. Expression occurred from late embryo to adult, especially in the pharynx. Other tissues, such as vulva and tail, had increased fluorescence during the late L3 and L4 stages. ife-4::GFP expression was not detected in germ line or somatic gonadal tissues other than spermatheca.
• Primary Identifier: Expr3459
• Reporter Gene: [ife-4::gfp] translational fusions. One was a simple array containing an NLS, ife-4::NLS::GFP(lsEx296), and the other, a complex array without an NLS, ife-4::GFP(lsEx385). The array with NLS was used to facilitate the identification of ife-4::GFP-expressing cells, whereas the array without NLS was expected to produce a subcellular distribution more similar to that of native IFE-4. ife-4::GFP was generated by using a PCR-based protocol. Briefly, the complete ife-4 coding sequence and 1.5 kb upstream of the 5$(B!l(B-trans-spliced acceptor site were amplified from genomic DNA (F, 5$(B!l(B-TGAAGCTTTCAATTTTCATTTCCAGGCG-3$(B!l(B; R, 5$(B!l(B-AGCTTGCATGCCTGCAGGTCGACTTTTGCAGATATTT-3$(B!l(B). The reverse primer contained a 24-nucleotide (nt) overlap to the GFP sequence. The GFP coding region, preceded by a nuclear localization signal (NLS) and followed by the unc-54 3$(B!l(B-UTR, was amplified from pPD95.67. Nested primers were used to create DNA encoding a fusion protein in a second PCR round. This DNA was microinjected into the gonad of N2 animals with 50 ng of the marker plasmid pRF4 carrying a dominant rol-6(su1006) allele to generate the extrachromosomal array lsEx296[ife-4::NLS::GFP rol-6(su1006)]. A second ife-4::GFP fusion was created as described above except that pPD95.67[Delta]NLS (the NLS was deleted by KpnI digestion and religation) was used in PCR. For microinjections, the second fusion product was mixed with linearized C. elegans genomic DNA to generate the complex extrachromosomal array lsEx385[ife-4::GFP rol-6(su1006)]. --precise ends.

9 Anatomy Terms

Definition	Name	Synonym	Primary Identifier
an accordion-like tube that contains sperm and is the site of oocyte fertilization.	spermatheca		WBbt:0005319
a large process bundle that runs along the vental mid-line extending from the ventral region of the nerve ring.	ventral nerve cord	ventral cord	WBbt:0005829
type of cells that make up muscle layers in the pharynx.	pharyngeal muscle cell		WBbt:0005451
muscle associated with hermaphrodite vulva.	vulval muscle		WBbt:0005821
neuron with its cell body situated in the tail, posterior to rectum.	tail neuron		WBbt:0006759
Neuron class of six motoneurons in the ventral cord, which innervate ventral body muscles and the vm2 muscles of the vulva.	VC neuron		WBbt:0005304
Herm. specific motor neurons (die in male embryo), innervate vulval muscles, serotonergic	HSNL	lineage name: ABplapppappa	WBbt:0004758
Herm. specific motor neurons (die in male embryo), innervate vulval muscles, serotonergic	HSNR	lineage name: ABprapppappa	WBbt:0004757
neuron of the pharyngeal nervous system.	pharyngeal neuron		WBbt:0005439

1 Genes

WormBase Gene ID	Gene Name	Sequence Name	Organism
WBGene00002062	ife-4	C05D9.5	Caenorhabditis elegans

6 Life Stages

Remark	Definition	Other Name	Public Name	Primary Identifier
	The stage that begins when a C.elegans individual is fully-developed and has reached maturity.		adult Ce	WBls:0000041
	The third stage larva. At 25 Centigrade, it ranges 32.5-40 hours after fertilization, 18.5-26 hours after hatch.		L3 larva Ce	WBls:0000035
	The second stage larva. At 25 Centigrade, it ranges 25.5-32.5 hours after fertilization, 11.5-18.5 hours after hatch.		L2 larva Ce	WBls:0000027
	The fourth stage larva. At 25 Centigrade, it ranges 40-49.5 hours after fertilization, 26-35.5 hours after hatch.		L4 larva Ce	WBls:0000038
	The first stage larva. At 25 Centigrade, it ranges 14-25.5 hours after fertilization, 0-11.5 hours after hatch.		L1 larva Ce	WBls:0000024
	The C. elegans life stage spanning 620-800min(hatch) after first cleavage at 20 Centigrade. Cell number remains at ~560 cells, with some new cells generated and some cells go through programmed cell death. A stage after elongation is over. The last stage of embryogenesis. Also called pre-hatched embryo, late embryo or morphogenetic embryo.		fully-elongated embryo Ce	WBls:0000021