WormBase Tree Display for Expr_pattern: Expr2632
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| Expr2632 | Expression_of | Gene | WBGene00002228 | |
|---|---|---|---|---|
| Reflects_endogenous_expression_of | WBGene00002228 | |||
| Expression_data | Life_stage | WBls:0000024 | ||
| WBls:0000003 | ||||
| WBls:0000038 | ||||
| WBls:0000027 | ||||
| WBls:0000035 | ||||
| WBls:0000041 | ||||
| Anatomy_term | WBbt:0004575 | Certain | ||
| WBbt:0004576 | Certain | |||
| WBbt:0005784 | Certain | |||
| GO_term | GO:0005938 | |||
| GO:0000922 | ||||
| GO:0005874 | ||||
| Subcellular_localization | During the meiotic divisions that occur in the male germ line, anti-KLP-18 staining patterns were similar to those seen in female meiosis and mitosis. KLP-18 was concentrated at the spindle poles, which have centrosomes, and in the anaphase interzone. There was also a significant amount of KLP-18 staining on male meiotic chromosomes. | |||
| Immunostaining showed KLP-18 colocalized with the disorganized MTs during prometaphase. As the MTs became ordered into a parallel bipolar array, KLP-18 concentrated most at the poles but was also present between the poles and chromosomes. The concentration at poles persisted during metaphase and early anaphase, but then shifted to the interzone between the separating chromosomes during late anaphase/telophase. Similar KLP-18 staining patterns were observed during meiosis II. | ||||
| KLP-18 staining was most concentrated in mitotic spindles toward the poles. During anaphase, KLP-18 staining also concentrated between the separating groups of chromosomes, with a dim zone at the spindle equator. After cytokinesis, KLP-18 accumulated somewhat around the nucleus and at the cortex near sites of cell-cell contact. Adult hermaphrodites contained high levels of cytoplasmic stain in the distal gonad arm. | ||||
| Type | Antibody | Polyclonal rabbit and rat antibody. PCR was used to amplify a 1.2 kb fragment of yk167g6, encoding the C-terminal 425 aa of KLP-18 (PARMA....TTKP): forward: 5-GAGGATCCCCTGCACGAATGGCTTTGTT-3, reverse: 5-ATGCGGTACCTGGCTTTGTGGTCTGACT-3. The fragment was cloned in frame into the 6xHis-tag expression vector pQE-30 (Qiagen), transformed, and induced in E. coli strain M15[pREP4] (Qiagen). Recombinant protein was purified on Ni-NTA matrix (Qiagen) and sent to Eurogentec for immunization. Final bleeds of one rabbit and one rat were purified using the 6xHis-KLP-18 fragment conjugated to AminoLink Plus (Pierce). | ||
| Pattern | During later stages of embryogenesis, KLP-18 levels gradually diminished in somatic cells but increased in the primordial germ cells Z2 and Z3, presumably due to expression in the embryonic germ line. The germ line stained brightly for KLP-18 at all stages of larval development. Adult hermaphrodites contained high levels of cytoplasmic stain in the distal gonad arm. Adults also showed high levels of staining in the proximal gonad arm. | |||
| Picture | WBPicture0000008730 | |||
| WBPicture0000008731 | ||||
| Remark | A dramatic reduction of staining in embryos after preincubation of the antibody with an excess of KLP-18 fusion protein. Staining was also greatly reduced or eliminated in embryos subjected to RNAi depletion of KLP-18. | |||
| Reference | WBPaper00006197 | |||
| Antibody_info | WBAntibody00000693 | |||
| WBAntibody00000694 |
