Fanconi anemia(FA)is a rare recessive hereditary disease characterized clinically by congenital defects,progressive bone-marrow failure,and cancer predisposition.Cells from FA patients exhibit hypersensitivity to DNA ...Fanconi anemia(FA)is a rare recessive hereditary disease characterized clinically by congenital defects,progressive bone-marrow failure,and cancer predisposition.Cells from FA patients exhibit hypersensitivity to DNA cross-linking agents,such as mitomycin C(MMC).To date,at least 12 FA genes have been found deleted or mutated in FA cells,and 10 FA gene products form a core complex involved in FA/BRCA2 DNA repair pathway-FA pathway.The ubiquitin E3 ligase FANCL,an important factor of FA core complex,co-functions with a new ubiquitin conjugating enzyme UBE2T to catalyze the monoubiquitination of FANCD2.FANCD2-Ub binds BRCA2 to form a new complex located in chromatin foci and then take part in DNA repair process.The deubiquitylating enzyme USP1 removes the mono-ubiquitin from FANCD2-Ub following completion of the repair process,then restores the blocked cell cycle to normal order by shutting off the FA pathway.In a word,the FANCD2 activity adjusted exquisitely by ubiquitination and/or deubiquitination in vivo may co-regulate the FA pathway involving in variant DNA repair pathway.展开更多
Gene targeting is a powerful approach of studying the gene function in vivo. Specific genetic modifications, including simple gene disruption, point mutations, large chromosomal deletions and rearrangements, targeted ...Gene targeting is a powerful approach of studying the gene function in vivo. Specific genetic modifications, including simple gene disruption, point mutations, large chromosomal deletions and rearrangements, targeted incorporation of foreign genes, could be introduced into the mouse genome by gene targeting. Recent studies make it possible to do the gene targeting with temporal and spatial control.展开更多
The human telomerase reverse transcriptase （hTERT） is expressed in more than 85% of tumor cells but is usually not found in normal cells, which makes hTERT as an ideal tumor-associate antigen （TAA） to develop pote...The human telomerase reverse transcriptase （hTERT） is expressed in more than 85% of tumor cells but is usually not found in normal cells, which makes hTERT as an ideal tumor-associate antigen （TAA） to develop potential vaccine specifically destroying cancers without impairing normal tissues in human cancer immunotherapy. Here are reviewed the fundamental advances of studies on immunogenicity of hTERT or its peptides and the early clinical trials using the hTERT vaccine approach in the last decades.展开更多
Western blot analysis revealed that one IgG<sub>1</sub> monoclonal antibody （mAb） to sp18 family membrane proteins （Mr. 14, 16 and 18 ku） of bovine sperm reacted faintly with protein bands of 14,18, 22...Western blot analysis revealed that one IgG<sub>1</sub> monoclonal antibody （mAb） to sp18 family membrane proteins （Mr. 14, 16 and 18 ku） of bovine sperm reacted faintly with protein bands of 14,18, 22, 30 and 60 ku （reducing） in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence （IIF） showed that the sp18 antigens were present in the posterior head of murine sperm. In murine in vitro fertilization （IVF） and embryo development trails, a total of 426 oocytes from C<sub>57</sub>BL/6 and F<sub>1</sub> hybrid strain (CD<sub>1</sub>×C<sub>57</sub>BL/6 cross) of 12, female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μg/mL of sp18 mAb in the modified TYH IVF medium for 15—20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 mAb groups was 77.1%, which was not significantly （P】0.05） different from the nonspecific mouse IgG（79.2 %） and non-IgG（80.3%） control groups. Fertilized oocytes had been展开更多
Conotoxins are short peptide-toxins with specific targets and large diversity. They are usetul in analgesia, neuroprotection, detection of some kinds of deseases, and receptor and ion channel study. in order to explor...Conotoxins are short peptide-toxins with specific targets and large diversity. They are usetul in analgesia, neuroprotection, detection of some kinds of deseases, and receptor and ion channel study. in order to explore the conotoxin resourses of Chinese oceans, rapid amplification of 3’ cDNA ends (RACE) method was utilized to systemically analyze the O-superfamily conotoxin content of Conus striatus inhabited near Chinese Hainan Island. Six new O-superfamily conopeptides were identified, one of which is highly homologous to MVIlA, an N-type calcium channel antagonist.展开更多
Transgenic mice with mammary gland secreting human granulocyte colony stimulating factor (G-CSF) were produced using mice whey acid protein gene promoter. It was found that there was very low expression level in mamma...Transgenic mice with mammary gland secreting human granulocyte colony stimulating factor (G-CSF) were produced using mice whey acid protein gene promoter. It was found that there was very low expression level in mammary gland. Human G-CSF cDNA was obtained by RT-PCR from transgenic mice mammary gland. Sequence analysis showed that this G-CSF gene deleted the 4th exon, and compared with human G-CSF genomic DNA, there were donor and acceptor splice sites in the deletion fragment. It was considered that the 3rd and 4th introns also delete in G-CSF fragment. The transgenic construct was corrected by deleting the 3rd and 4th introns to construct the minigene, which was used to produce transgenic mice by microinjection. Northern blot showed that G-CSF expression using the new construct increased 5.4 times as that before in transgenic mice. The results suggested that it was possible that RNA aberrant splice result in low expression in transgenic mice.展开更多
基金was supported by the National Natural Sciences Foundation of China(No.30470379).
文摘Fanconi anemia(FA)is a rare recessive hereditary disease characterized clinically by congenital defects,progressive bone-marrow failure,and cancer predisposition.Cells from FA patients exhibit hypersensitivity to DNA cross-linking agents,such as mitomycin C(MMC).To date,at least 12 FA genes have been found deleted or mutated in FA cells,and 10 FA gene products form a core complex involved in FA/BRCA2 DNA repair pathway-FA pathway.The ubiquitin E3 ligase FANCL,an important factor of FA core complex,co-functions with a new ubiquitin conjugating enzyme UBE2T to catalyze the monoubiquitination of FANCD2.FANCD2-Ub binds BRCA2 to form a new complex located in chromatin foci and then take part in DNA repair process.The deubiquitylating enzyme USP1 removes the mono-ubiquitin from FANCD2-Ub following completion of the repair process,then restores the blocked cell cycle to normal order by shutting off the FA pathway.In a word,the FANCD2 activity adjusted exquisitely by ubiquitination and/or deubiquitination in vivo may co-regulate the FA pathway involving in variant DNA repair pathway.
基金the National Natural Science Foundation of China (Grant Nos. 39970413 and 30070837) the National High Technology Research and Development Program (Grant No. 102-08-08-02) Beijing Science Projects (Grant No. 954020600) and the Innovation Initiation Fu
文摘Gene targeting is a powerful approach of studying the gene function in vivo. Specific genetic modifications, including simple gene disruption, point mutations, large chromosomal deletions and rearrangements, targeted incorporation of foreign genes, could be introduced into the mouse genome by gene targeting. Recent studies make it possible to do the gene targeting with temporal and spatial control.
文摘The human telomerase reverse transcriptase （hTERT） is expressed in more than 85% of tumor cells but is usually not found in normal cells, which makes hTERT as an ideal tumor-associate antigen （TAA） to develop potential vaccine specifically destroying cancers without impairing normal tissues in human cancer immunotherapy. Here are reviewed the fundamental advances of studies on immunogenicity of hTERT or its peptides and the early clinical trials using the hTERT vaccine approach in the last decades.
文摘Western blot analysis revealed that one IgG<sub>1</sub> monoclonal antibody （mAb） to sp18 family membrane proteins （Mr. 14, 16 and 18 ku） of bovine sperm reacted faintly with protein bands of 14,18, 22, 30 and 60 ku （reducing） in samples of mouse sperm. The mAb also reacted to protein of egg lysozyme. Using a laser confocal microscope, indirect immunofluorescence （IIF） showed that the sp18 antigens were present in the posterior head of murine sperm. In murine in vitro fertilization （IVF） and embryo development trails, a total of 426 oocytes from C<sub>57</sub>BL/6 and F<sub>1</sub> hybrid strain (CD<sub>1</sub>×C<sub>57</sub>BL/6 cross) of 12, female mice were used in 3 independent trails. After preincubating capacitated sperm with 182 μg/mL of sp18 mAb in the modified TYH IVF medium for 15—20 min, cumulus-oocyte-complexes were introduced. The fertilization rate in sp18 mAb groups was 77.1%, which was not significantly （P】0.05） different from the nonspecific mouse IgG（79.2 %） and non-IgG（80.3%） control groups. Fertilized oocytes had been
文摘Conotoxins are short peptide-toxins with specific targets and large diversity. They are usetul in analgesia, neuroprotection, detection of some kinds of deseases, and receptor and ion channel study. in order to explore the conotoxin resourses of Chinese oceans, rapid amplification of 3’ cDNA ends (RACE) method was utilized to systemically analyze the O-superfamily conotoxin content of Conus striatus inhabited near Chinese Hainan Island. Six new O-superfamily conopeptides were identified, one of which is highly homologous to MVIlA, an N-type calcium channel antagonist.
文摘Transgenic mice with mammary gland secreting human granulocyte colony stimulating factor (G-CSF) were produced using mice whey acid protein gene promoter. It was found that there was very low expression level in mammary gland. Human G-CSF cDNA was obtained by RT-PCR from transgenic mice mammary gland. Sequence analysis showed that this G-CSF gene deleted the 4th exon, and compared with human G-CSF genomic DNA, there were donor and acceptor splice sites in the deletion fragment. It was considered that the 3rd and 4th introns also delete in G-CSF fragment. The transgenic construct was corrected by deleting the 3rd and 4th introns to construct the minigene, which was used to produce transgenic mice by microinjection. Northern blot showed that G-CSF expression using the new construct increased 5.4 times as that before in transgenic mice. The results suggested that it was possible that RNA aberrant splice result in low expression in transgenic mice.
