Rs. C3GSH3-b domain was amplified by PCR with oligos > 자유게시판

Rs. C3GSH3-b domain was amplified by PCR with oligos C3GSH3-bF: 5 GGGAATTCCCATGGCTGGCATTCGGGTGGTTGAT-3?and C3GSH3-bR: 5 CCGGATCCCTACTAACTGCCGTCTCTGCTGTCC-3?and cloned into NcoIBamHI sites of pETEV15b. Cbl-SH3-b domain was amplified with oligos CblSH3bBamHI-F:5?GGGGGATCCCCGCCTTCTCCATTCTC-3?and CblSH3bXhoI-R: 5?CCCCTCGAGCTACTAAGGTGGCAGTTTTGGCAC-3?and cloned into pET15b-NBKSXa by BamHI-XhoI digestion. p130CasP2-domain (proline-rich region 2) was amplified with oligos CasP2BamHI-F: 5 GGGGATCCTCACTGCTCTTCAGACGG-3 and CasP2XhoI-R: 5 GGCTCGAGCTACTAGGTGAACTTAGGGGGTGA-3?and cloned into BamHIXhoI sites of pET15b-NBKSXa. Competing interests The authors declare that they have no competing interests. Authors' contributions VM carried out the pull-down experiments, VM and SO-R made the 2-Bromo-1,3-difluoro-4-nitrobenzene immunofluorescences, immunoprecipitations and PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/12958591 cell adhesion assays, MS and JG-B made all the constructs, VM and MS performed the shRNA lentiviral constructs, VM and IF-A participated in the SH3 arrays hybridizations, VM and SG-H performed the immunoblottings, CG, JMP and AP designed the experiments, CG wrote the paper, AP and JMP critically revised the paper. All authors read and approved the final manuscript. Acknowledgements This work was supported by grants form the Spanish Ministry of Health (ISCIII) [FIS-PI070078 to CG and FIS-PI070071 to AP], by grants from the Spanish Ministry of Economy and Competitiveness [SAF2010-20918-C02-02 to CG, SAF2010-20918-C02-01 to AP and BFU2009-08389 to JMP], by a grant for Research in Biomedicine from the Council of Health and Social Welfare of Junta de Castilla y Le and by grants from the Council of Education 3-Bromo-4-(propan-2-yloxy)aniline hydrochloride of Junta de Castilla y Le [CSI06A09 and SA157A12-1], Spain (to JMP and CG). All funding was cosponsored by the European FEDER Program. Author details 1 Centro de Investigaci PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25386826 del C cer, IBMCC, CSIC-Universidad de Salamanca, Salamanca, Spain. 2Departamento de Bioqu ica y Biolog Molecular II, Facultad de Farmacia, UCM, Instituto de Investigaci Sanitaria del Hospital Cl ico San Carlos (IdISSC), Madrid, Spain. 3Departamento de Medicina, Facultad de Medicina, Universidad de Salamanca; Instituto de Investigaci Biom ica de Salamanca (IBSAL), Salamanca, Spain. Received: 31 July 2012 Accepted: 18 January 2013 Published: 23 January3.4.5.6.7.8.9.10.11. 12.13.14.15.16.17.18.19. 20.21. References 1. Birge RB, Kalodimos C, Inagaki F, Tanaka S: Crk and CrkL adaptor proteins: networks for physiological and pathological signaling. Cell Commun Signal 2009, 7:13. 2. Shivakrupa R, Radha V, Sudhakar C, Swarup G: Physical and functional interaction between Hck tyrosine kinase and guanine nucleotide exchange factor C3G results in apoptosis, which is independent of C3G catalytic domain. J Biol Chem 2003, 278:52188?2194.22.23.Radha V, Rajanna A, Mitra A, Rangaraj N, Swarup G: C3G is required for cAbl-induced filopodia and its overexpression promotes filopodia formation. Exp Cell Res 2007, 313:2476?492. Ohba Y, Ikuta K, Ogura A, Matsuda J, Mochizuki N, Nagashima K, Kurokawa K, Mayer BJ, Maki K, Miyazaki J, Matsuda M: Requirement for C3Gdependent Rap1 activation for cell adhesion and embryogenesis. EMBO J 2001, 20:3333?341. Radha V, Mitra A, Dayma K, Sasikumar K: Signalling to actin: role of C3G, a multitasking guanine-nucleotide-exchange factor. Biosci Rep 2011, 31:231?44. Arai A, Nosaka Y, Kohsaka H, Miyasaka N, Miura O: CrkL activates integrinmediated hematopoietic cell adhesion through the guanine nucleotide exchange factor C3G. Blood 1999, 93.