Plasmid > pgat-HW26

pgat-HW26

Xenbase Number: n/a
Previous Plasmid name: pJC2LXG1
Species: Xla
Gene symbol: pgat
Gene Name: Primordial germ cell-associated transcript protein
Gene synonyms: xpat
Gene Function: germplasm formation
Vector: pSPJC2L
Antibiotic: Amp
Purpose: To generate mRNA for injection
Special Features: A GFP fusion with GFP at the C-terminus. Xpat-GFP for micro-injection was made by inserting the PCR Amplified ORF of GFP (primers: 5’gccgagatctATGAGTAAAGGAGAAGAAC 3’; 5’ gccggaattcTTATTTGTATAGTTCATCCAT 3’) between the BglII and EcoR1 sites of pSPJC2L(Cook et al., 1993) to give pSPJC2L-cGFP. TheORF of Xpat without the STOP codon was Amplified by PCR (primers: 5’gccgagatctATGGCTTTGAAGGCAGAAG 3’;5’gccgagatctCAGGCCAACATCCCACCAAG 3’) and inserted into the BglII site of pSPJC2L-cGFP to yield pSPJC2 Xpat GFP. Sense RNA was transcribed from this NotI-linearized plasmid with SP6 RNA polymerase. This clone forms germ plasm-like islands in oocytes under very specic culture conditions
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Sense Linearization: NotI
Sense Polymerase: SP6
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Reference: Machado, R. J., Moore, W., Hames, R., Houliston, E., Chang, P., King, M. L., Woodland, H. R., 2005. Xenopus Xpat protein is a major component of germ plasm and may function in its organisation and positioning. Developmental Biology. 287, 289-300 Hudson, C. and Woodland, H. R (1998) 'Xpat, a gene expressed specifically in germ plasm and primordial germ cells of Xenopus laevis', Mechanisms Of Development, 73 159 - 168 (0925-4773)
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