RÉSUMÉ
<p><b>OBJECTIVE</b>Construct the gene library of apoptosis related genes in acute promyelocytic leukemia (APL) cell line NB4 cells treated by arsenic trioxide to clarify the apoptotic mechanism of NB4 cells.</p><p><b>METHOD</b>APL cell line NB4 cells treated with or without arsenic trioxide for 24 hours. Total RNA was extracted and suppress subtractive hybridization (SSH) was conducted according to the manual. With the cDNA of the apoptosis cells as the tester and that of control cells as the driver, forward and reverse hybridization was performed. Differentially expressed genes were linked with pGEM-Teasy cloning vector and transformed into E. coli DH5alpha. The positive clones were screened by blue and white spot. PCR were used to amplify these genes.</p><p><b>RESULT</b>The subtractive cDNA libraries related with apoptosis of NB4 cells were successfully constructed.</p><p><b>CONCLUSION</b>The constructed subtractive libraries are suitable for further study on the functional genes associated with apoptosis ofNB4 cells induced by arsenic trioxide.</p>
Sujet(s)
Humains , Apoptose , Génétique , Composés de l'arsenic , Pharmacologie , Lignée cellulaire tumorale , ADN complémentaire , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes tumoraux , Banque de gènes , Leucémie aiguë promyélocytaire , Anatomopathologie , Hybridation d'acides nucléiques , Méthodes , Séquençage par oligonucléotides en batterie , Oxydes , Pharmacologie , Analyse de séquence d'ADNRÉSUMÉ
The mutation of human cereblon gene (CRBN) is revealed to be related with mild mental retardation. Since the molecular characteristics of CRBN have not been well presented, we investigated the general properties of CRBN. We analyzed its gene structure and protein homologues. The CRBN protein might belong to a family of adenosine triphosphate (ATP)-dependent Lon protease. We also found that CRBN was widely expressed in different tissues, and the expression level in testis is significantly higher than other tissues. This may suggested it could play some important roles in several other tissues besides brain. Transient transfection experiment in AD 293 cell lines suggested that both CRBN and CRBN mutant (nucleotide position 1,274(C > T)) are located in the whole cells. This may suggest new functions of CRBN in cell nucleolus besides its mitochondria protease activity in cytoplasm.
Sujet(s)
Déficience intellectuelle/génétique , Peptide hydrolases/génétique , Protéines adaptatrices de la transduction du signal , Séquence d'acides aminés , Lignée cellulaire , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes , Humains , Données de séquences moléculaires , Peptide hydrolases/composition chimique , Transport des protéines , ARN messager/génétique , ARN messager/métabolisme , RT-PCR , Alignement de séquences , Analyse de séquence d'ADN , Fractions subcellulaires/métabolisme , Ubiquitin-protein ligasesRÉSUMÉ
Objective To study the effect of-1 site single nucleotide polymorphism(SNP) on CCNH gene promoter transcription activity.Methods PCR and site-directed mutagenesis technology were used to construct CCNH basic promoter and-1G mutate promoter.Dual-Luciferase Reporter assay system was used to detect the transcription activity of constructed promoter.Results In AD293 cells,the activity of-1G mutate type promoter was significantly lower than that of wild type-1T promoter(P
RÉSUMÉ
NF-kappaB-repression factor (NRF) is a nuclear inhibitor of NF-kappaB proteins that can silence the IFNbeta promoter. Since NRF was cloned in 1999, in-depth studies have been conducted on the biological functions of this constitutive repressor of NF-kappaB proteins. During large-scale sequencing of a human fetal brain cDNA library we isolated a novel human cDNA that proved to be a correct full-length NRF cDNA. The deduced protein contains 690 aa, and has a G-patch and an R3H domain at its C-terminus. The size of the protein is consistent with its counterparts in mouse and rat. There is considerable evidence that there are some mistakes in the NRF cDNA sequence reported by Nourbakhsh. Here we report the correct, full-length cDNA and protein sequences of NRF. Full-length NRF cDNA is 3247 bp long, contains three exons and maps to human chromosome Xq24. RT-PCR shows that NRF is widely expressed in human tissues.
Sujet(s)
Protéines de liaison à l'ADN/génétique , Facteur de transcription NF-kappa B/antagonistes et inhibiteurs , Protéines de répression/génétique , Facteurs de transcription/génétique , Séquence d'acides aminés , Séquence nucléotidique , Clonage moléculaire , ADN complémentaire , Protéines de liaison à l'ADN/métabolisme , Humains , Données de séquences moléculaires , Protéines de répression/métabolisme , Analyse de séquence d'ADN , Facteurs de transcription/métabolismeRÉSUMÉ
<p><b>OBJECTIVE</b>To explore the distribution of monoamine oxidase A (MOA-A) EcoRV polymorphism in Shanghai Han population and its possible role in the risk for Parkinson's disease(PD).</p><p><b>METHODS</b>The MAO-A gene EcoRV polymorphism was detected with PCR-RFLP method in 110 PD patients and 182 healthy controls, furthermore, statistical analysis was performed to investigate association between EcoR V polymorphism and PD onset.</p><p><b>RESULTS</b>(1)Remarkable difference in MAO-A EcoR V polymorphic distribution has been observed between Shanghai Han population and that in North America. (2) Neither allelic frequency nor genotypic frequency in PD cases differs significantly from that in healthy controls regardless of data from male or female subclass.</p><p><b>CONCLUSION</b>There may be racial difference in the distribution of the human MAO-A EcoR V (C/T) polymorphism, but the present research does not support the association between this variant and susceptibility to PD in Chinese Han population of Shanghai area.</p>
