The Experts below are selected from a list of 75 Experts worldwide ranked by ideXlab platform
Jianping Xiao - One of the best experts on this subject based on the ideXlab platform.
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characterization of an extremely thermostable restriction enzyme pspgi from a pyrococcus strain and cloning of the pspgi restriction modification system in escherichia coli
Applied and Environmental Microbiology, 1998Co-Authors: Richard D Morgan, Jianping XiaoAbstract:An extremely thermostable restriction endonuclease, PspGI, was purified from Pyrococcus sp. strain GI-H. PspGI is an isoschizomer of EcoRII and cleaves DNA before the first C in the sequence 5' CCWGG 3' (W is A or T). PspGI digestion can be carried out at 65 to 85 degrees C. To express PspGI at high levels, the PspGI restriction-modification genes (pspGIR and pspGIM) were cloned in Escherichia coli. M.PspGI contains the conserved sequence motifs of alpha-Aminomethyltransferases; therefore, it must be an N4-cytosine methylase. M.PspGI shows 53% similarity to (44% identity with) its isoschizomer, M.MvaI from Micrococcus variabilis. In a segment of 87 amino acid residues, PspGI shows significant sequence similarity to EcoRII and to regions of SsoII and StyD4I which have a closely related recognition sequence (5' CCNGG 3'). PspGI was expressed in E. coli via a T7 expression system. Recombinant PspGI was purified to near homogeneity and had a half-life of 2 h at 95 degrees C. PspGI remained active following 30 cycles of thermocycling; thus, it can be used in DNA-based diagnostic applications.
Jennifer R Toone - One of the best experts on this subject based on the ideXlab platform.
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recurrent mutations in p and t proteins of the glycine cleavage complex and a novel t protein mutation n145i a strategy for the molecular investigation of patients with nonketotic hyperglycinemia nkh
Molecular Genetics and Metabolism, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:Abstract Screening a DNA bank from 50 patients with enzymatic confirmation of their diagnosis of nonketotic hyperglycinemia gave allele frequencies of 5% for R515S of P-protein (glycine decarboxylase) and 7% for R320H of T-protein (Aminomethyltransferase). In a previous report we found that 3% of the same patient alleles were positive for T-protein IVS7−1G>A. In total, testing for these three mutations identified 15% of alleles and positive results (one or two mutations) were found in 11 of the 50 patients. In addition, a novel point mutation in T-protein, N145I, was found in a single case and a PCR/restriction enzyme assay was developed for its detection.
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identification of the first reported splice site mutation ivs7 1g a in the Aminomethyltransferase t protein gene amt of the glycine cleavage complex in 3 unrelated families with nonketotic hyperglycinemia
Human Mutation, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:A novel splice site mutation (IVS7-1G-->A) in the T-protein gene (Aminomethyltransferase, or AMT) of the glycine cleavage enzyme complex was found in a patient with nonketotic hyperglycinemia (NKH). A PCR/restriction enzyme method to detect this mutation was used to screen 100 NKH alleles and identified the mutation in three unrelated families.
Erick R James - One of the best experts on this subject based on the ideXlab platform.
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recurrent mutations in p and t proteins of the glycine cleavage complex and a novel t protein mutation n145i a strategy for the molecular investigation of patients with nonketotic hyperglycinemia nkh
Molecular Genetics and Metabolism, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:Abstract Screening a DNA bank from 50 patients with enzymatic confirmation of their diagnosis of nonketotic hyperglycinemia gave allele frequencies of 5% for R515S of P-protein (glycine decarboxylase) and 7% for R320H of T-protein (Aminomethyltransferase). In a previous report we found that 3% of the same patient alleles were positive for T-protein IVS7−1G>A. In total, testing for these three mutations identified 15% of alleles and positive results (one or two mutations) were found in 11 of the 50 patients. In addition, a novel point mutation in T-protein, N145I, was found in a single case and a PCR/restriction enzyme assay was developed for its detection.
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identification of the first reported splice site mutation ivs7 1g a in the Aminomethyltransferase t protein gene amt of the glycine cleavage complex in 3 unrelated families with nonketotic hyperglycinemia
Human Mutation, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:A novel splice site mutation (IVS7-1G-->A) in the T-protein gene (Aminomethyltransferase, or AMT) of the glycine cleavage enzyme complex was found in a patient with nonketotic hyperglycinemia (NKH). A PCR/restriction enzyme method to detect this mutation was used to screen 100 NKH alleles and identified the mutation in three unrelated families.
Richard D Morgan - One of the best experts on this subject based on the ideXlab platform.
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characterization of an extremely thermostable restriction enzyme pspgi from a pyrococcus strain and cloning of the pspgi restriction modification system in escherichia coli
Applied and Environmental Microbiology, 1998Co-Authors: Richard D Morgan, Jianping XiaoAbstract:An extremely thermostable restriction endonuclease, PspGI, was purified from Pyrococcus sp. strain GI-H. PspGI is an isoschizomer of EcoRII and cleaves DNA before the first C in the sequence 5' CCWGG 3' (W is A or T). PspGI digestion can be carried out at 65 to 85 degrees C. To express PspGI at high levels, the PspGI restriction-modification genes (pspGIR and pspGIM) were cloned in Escherichia coli. M.PspGI contains the conserved sequence motifs of alpha-Aminomethyltransferases; therefore, it must be an N4-cytosine methylase. M.PspGI shows 53% similarity to (44% identity with) its isoschizomer, M.MvaI from Micrococcus variabilis. In a segment of 87 amino acid residues, PspGI shows significant sequence similarity to EcoRII and to regions of SsoII and StyD4I which have a closely related recognition sequence (5' CCNGG 3'). PspGI was expressed in E. coli via a T7 expression system. Recombinant PspGI was purified to near homogeneity and had a half-life of 2 h at 95 degrees C. PspGI remained active following 30 cycles of thermocycling; thus, it can be used in DNA-based diagnostic applications.
Derek A Applegarth - One of the best experts on this subject based on the ideXlab platform.
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recurrent mutations in p and t proteins of the glycine cleavage complex and a novel t protein mutation n145i a strategy for the molecular investigation of patients with nonketotic hyperglycinemia nkh
Molecular Genetics and Metabolism, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:Abstract Screening a DNA bank from 50 patients with enzymatic confirmation of their diagnosis of nonketotic hyperglycinemia gave allele frequencies of 5% for R515S of P-protein (glycine decarboxylase) and 7% for R320H of T-protein (Aminomethyltransferase). In a previous report we found that 3% of the same patient alleles were positive for T-protein IVS7−1G>A. In total, testing for these three mutations identified 15% of alleles and positive results (one or two mutations) were found in 11 of the 50 patients. In addition, a novel point mutation in T-protein, N145I, was found in a single case and a PCR/restriction enzyme assay was developed for its detection.
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identification of the first reported splice site mutation ivs7 1g a in the Aminomethyltransferase t protein gene amt of the glycine cleavage complex in 3 unrelated families with nonketotic hyperglycinemia
Human Mutation, 2001Co-Authors: Jennifer R Toone, Derek A Applegarth, Marion B Coultermackie, Erick R JamesAbstract:A novel splice site mutation (IVS7-1G-->A) in the T-protein gene (Aminomethyltransferase, or AMT) of the glycine cleavage enzyme complex was found in a patient with nonketotic hyperglycinemia (NKH). A PCR/restriction enzyme method to detect this mutation was used to screen 100 NKH alleles and identified the mutation in three unrelated families.