For the detection of Bmp2, a 460 bp fragment encoding the divergent propeptide domain of BMP2 was used (Andrée et al., 1998), generated by RT-PCR using primers 5′-TAATGAGGAGTCTGTCACC-3′ and 5′-TGTATTTGTGGCGCTTACGC-3′
Authors indicated the probe cDNA was provided by Dr. P. Brickell (Francis et al., ), who indicated "The longest chicken Bmp-2 cDNA clone had an insert of 1120 bp ... (and they used)
nucleotides 1-797 ... as a template." Although the full length BMP2 gene is 1179 nt and thus the probe cannot be precisely known, we took the first 797 of the full length clone to be this probe.
The following probes were used: Sf/Hgf (Thery et al., 1995), qPax3, MyoD and Myf5, Lbx1, Bmp2 and Bmp4, Tcf4. Probes used in this study were kindly provided by Claudio Stern, Gabrielle Kardon, Christophe Marcelle, Bruce M. Paterson, Susanne Dietrich and Paul Brickell. Thus insufficient information provided in publication to verify exact sequence used to synthesize probe. The sequence below was obtained from NCBI (acc # NM_204358).
DIG-labeled probes were generated for Gremlin (Capdevila et al., 1999), Shh (Riddle et al., 1993), Bmp2 (Francis et al., 1994)... Francis et al reported: "The longest chicken Bmp-2 cDNA clone had an insert of 1120 bp
and was designated p5. ... To construct a template for the synthesis of Bmp-2-specific probes,
nucleotides 1-797 of p5 were amplified by PCR." Although the full length clone is 1179nt, so the exact probe cannot be determined, the first 797nt of NM_204358 are included here as the putative probe.
Insufficient information provided in publication to verify exact sequence used to synthesize probe. Sequence below was obtained from NCBI (acc # NM_204358). Images are quail, but sequence and map shown are chick.
Authors indicated the probe sequence was from the clone identified in Francis-West et al., 1994. This clone has NCBI acc# NM_204358.1. This paper said:
nucleotides 1-797 (were)cloned into pBluescript SK+ ...Antisense RNA probes specific for
chicken Bmp-2 transcripts were synthesized with T3 RNA polymerase,
using HindIII-linearized p5.1 as a template.
Copyright Vargesson, Laufer. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
According to the authors, "Riboprobes cBmp2, cBmp4 and cBmp7 were kindly provided by C. Tabin and B. Houston." As information provided was insufficient to verify exact sequence used to synthesize probe, the sequence was obtained from NCBI (acc# NM_204358.1).
Images provided by Claire Anderson. The publication of Liem et al. 1995 (Liem Jr., K.F, Tremml G, Roelink H, Jessell T.M. Dorsal differentiation of neural plate cells induced by BMP-mediated signals from epidermal ectoderm. Cell. 1995; 82: 969-979. PMID: 7553857 ) was referred for the probe information. Authors indicated further probe information could be found in another publication (Francis PH, Richardson MK, Brickell PM, Tickle C. Bone morphogenetic proteins and a signalling pathway that controls patterning in the developing chick limb. Development. 1994 Jan;120(1):209-18. PMID: 8119128). The indicated probe sequence was extracted from NCBI (GenBank: X75914.1).