A novel mutation of NTRK1 gene in a family with congenital insensitivity to pain with anhidrosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To screen for mutations in the neurotrophic tyrosine kinase receptor type 1 (NTRK1) gene in a Chinese family affected with congenital insensitivity to pain with anhidrosis (CIPA).</p><p><b>METHODS</b>With informed consent obtained, peripheral blood samples were obtained from the patient and his family members. Seventeen coding exons and intron-exon boundaries of the NTRK1 gene were amplified with PCR and analyzed by direct sequencing.</p><p><b>RESULTS</b>A novel mutation c.2086_2087insC (p.Arg696 fsx) was identified in exon 16 of the NTRK1 gene in the proband. This insertion has caused open reading frame shifting and a premature termination has occurred just one codon downstream. Truncation of 72 amino acids at the C terminus has wiped out part of the tyrosine kinase domain (TKD) of the protein. Both of the proband's parents and two grandmothers have carried the c.2086_2087insC (p.Arg696 fsx) mutation. No mutation was found in the NTRK1 gene of other siblings.</p><p><b>CONCLUSION</b>Mutation analysis of the NTRK1 gene has been carried out in a Chinese family affected with CIPA, and a novel NTRK1 gene mutation was identified.</p>

Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli / 中华男科学杂志

<p><b>OBJECTIVES</b>To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21.</p><p><b>METHODS</b>The coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers, and cloned into pGEX-2T after the restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG.</p><p><b>RESULTS</b>Sequencing and restriction digestion of the recombinant plasmid revealed the existence of coding sequence for mouse lactate dehydrogenase subunit C. A protein band of about 60,000 could be induced by IPTG in the recombinant plasmid.</p><p><b>CONCLUSIONS</b>The coding sequence of mouse lactate dehydrogenase subunit C was introduced into the pGEX-2T plasmid and a GST-fused protein could be induced at a high level.</p>

Method for molecular diagnosis of hereditary methemoglobinemia / 中华检验医学杂志

Objective To investigate molecular diagnostic method for hereditarymethemoblobinemia. Methods The cDNA coding sequence of NADH-cytochrome b5 reductase (b5R) from 3 patients with hereditary methemoglobinemia was analyzed by direct sequencing of RT-PCR products and the genomic DNA of b5R gene by PCR-restriction endonuclease digestion or PCR-sequencing. Results The b5R cDNA of patient A was T/C heterozygous at nucleotide 527 and G/A heterozygous at nucleotide 608. The b5R cDNA of patient B was G/A heterozygous at both nucleotide 170 and nucleotide 179. The b5R cDNA of patient C was G/A heterozygous at nucleotide 608 and C/T heterozygous at nucleotide 791. Result of genomic DNA analysis was in agreement with that of cDNA approach. Conclusion The method for molecular diagnosis of hereditary methemoglobinemia was established and 3 novel b5R gene mutations were identified in compound heterozygosity in 3 Chinese patients.

Gene-based individual identification of hardly recognizable victims in an accident / 解放军医学杂志

Objective To identify hardly recognizable victims of an accident. Methods Tissues of muscle and cartilage were obtained from the dead bodies. Some of the tissues were checked by routine pathological microscopy. Genomic DNA was isolated from the tissues and subjected to STR profiling of 16 sites via multiple fluorescent PCR analysis with ABI’s AmpFLSTR Amplification Kit. Individual identification of the victims was carries out by matching the STR profiles of the victims with those of the parents. Results Routine pathological microscopy showed that the structure of some of the muscle tissues was totally destroyed, while the structure of all cartilage tissues was basically intact. Three patterns of genomic DNA isolated from victims’ muscle tissues could be seen in gel electrophoresis, i.e. basically undegraded, partially degraded and totally degraded. STR profiling failed due to the degradation of genomic DNA of some of the muscle tissues, while all samples of the cartilage genomic DNA could be used for STR typing. Conclusion Paternity identification based on STR genotyping was an effective way to identify victims of accidents, and cartilage tissue from the victims was the first choice for that purpose.