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71.
72.
CAO Cong ZHAO GuoWei YU Wei XIE XueMin WANG WenTian YANG RuiFeng LV Xiang LIU DePei 《中国科学:生命科学英文版》2014,57(5):488-494
Studies on the chaperone proteinα-hemoglobin stabilizing protein(AHSP)reveal that abundant AHSP in erythroid cells enhance the cells’tolerance to oxidative stress imposed by excessα-hemoglobin in pathological conditions.However,the potential intracellular modulation of AHSP expression itself in response to oxidative stress is still unknown.The present study examined the effect and molecular mechanism of STAT3,an oxidative regulator,on the expression of AHSP.AHSP expression increased in K562 cells upon cytokine IL-6-induced STAT3 activation and decreased in STAT3 knock-down K562 cells.Regulation of AHSP in oxidative circumstance was then examined inα-globin-overloaded K562 cells,and real-time PCR showed strengthened expression of both AHSP and STAT3.ChIP analysis showed binding of STAT3 to AHSP promoter and binding was significantly augmented with IL6 stimulation and uponα-globin overexpression.Dual luciferase reporter assays of the wildtype and mutated SB3 element,an IL-6RE site,in the AHSP promoter in K562 cells highlighted the direct regulatory effect of STAT3 on AHSP gene.Finally,direct binding of STAT3 to SB3 site of AHSP promoter was confirmed with EMSA assays.Our work reveals an adaptive AHSP regulation mediated by the redox-sensitive STAT3 signaling pathway,and provides clues to the therapeutic strategy for AHSP enhancement. 相似文献
73.
展望21世纪的生命科学 总被引:17,自引:0,他引:17
对本世纪以来发展迅猛异常的生物科学的概况,特别是近对二三十年来,在分子生物学的带动下,生命科学在结构分子生物学;中心法则及其后的生物学细节研究;基因表达、调控和发育生物学;基因组计划和应用生物学及生物技术的发展;宏观生物学等方面讨论了生命科学的发展和特点,并从四个主要方面对21世纪生命科学(包括生物技术)的发展趋势进行了预测,以达到综览生命科学全局,明确今后发展方向的目的。 相似文献
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75.
73例中国人血友病甲基因突变的分析 总被引:5,自引:1,他引:5
刘建湘 张宇舟 王鸿利 黄秋花 曹文俊 王学锋 邵慧珍 王振义 陈竺 黄薇LIU Jian-Xiang ZHANG Yu-Zhou WANG Hong-Li HUANG Qiu-Hua CAO Wen-Jun WANG Xue-Feng SHAO Hui-Zhen WANG Zhen-Yi CHEN Zhu HANG Wei 《遗传》1998,20(1):1-6
我们用Southern blotting、PCR、变性梯度凝胶电泳(DGGE)和DNA测序等方法对73例血友病甲患者(经上海瑞金医院测定血浆FVⅢ:C和vWF:Ag诊断,其中无亲缘关系患者65例,按FVⅢ:C水平分为轻、中、重三型。FVⅢ:C< 2%为重型,共47例;FVⅢ:C 2%-5%为中型,共9例;FVⅢ:C5%-25%为轻型,共1 7例)进行FVⅢ基因突变检测。共检出内含子22倒位23例,均为重型,约占重型的49%,与国外报道相似。余下50例(其中无亲缘关系者45)用PCR-DGG E分析所有外显子及其侧翼内含子序列,发现异常条带则进行DNA测序。在17例患者中检出突变13种,其中无义突变5种,均为重型;错义突变6种,除1例外都是轻中型;小缺失2例,都是重型;其中,AA466Lys(AAG)-Thr(ACG)、719Tyr(TAC)-Stop(TAG)、AA826 Asp(GAC)-Glu(GAA)、312Ile(ATC)-xxC及AA1551-1552del(AGAA)为新发现的突变。有亲缘关系的患者都有相同的基因突变,而在无亲缘关系患者未发现相同突变。基因突变与临床表现基本相符。
Abstract:We use Southern blotting,PCR,denaturing gradient gel electrophoresis(DGGE)and DNA sequencing to detect gene mutations of haemophilia A in Chinese population.73 cases(47 severe)(FVIII:C<2%),9 moderate(FVIII:C 2%~5%),17 mild(FVIII:C 5%~25%)of haemophilia A were first screened with Southern blotting,23 were found to be the intron 22 inversion type,all being severe cases.The remaining 50 cases without intron 22 in version were examined with PCR-DGGE.Genomic DNA were amplified using GC-clamped primers covering all the exons and all flanking intron regions.Abnormal bands were sequenced.13 different mutations were identified,including 5 nonsense mutations,6 missense mutations and 2 small deletions.5 mutations,AA466Lys(AAG)-Thr(ACG),AA719Tyr(TAC)-Stop(TAG),AA826Asp(GAC)-Glu(GAA),AA312Ile(ATC)-xxC and AA1551-1552del(AGAA)have not been reported before.Generally the genetic defects correspond to the clinical conditions. 相似文献
76.
马铃薯试管薯诱导及其遗传转化体系的优化 总被引:5,自引:0,他引:5
目的:对马铃薯试管薯诱导及遗传转化体系进行优化研究。方法:利用3种培养法对马铃薯品种甘农薯2号和Favorita进行了试管薯的诱导,并用农杆菌介导法对其遗传转化体系进行优化研究。结果:用固体培养、固液培养和液体培养法诱导的甘农薯2号试管薯单瓶平均结薯数分别为4.6、4.4和6.5个,块茎平均直径分别为6.2、5.8和7.7ram;而Favorita单瓶平均结薯数为5.4、5.8和7.4个,平均直径分别达6.2、5.9和7.3mm。用浓度为OD600=0.5的农杆菌菌液侵染8min,共培养2d能够获得较高的转化频率。结论:液体培养法诱导试管薯的效果最好,建立的高频率遗传转化体系使甘农薯2号和Favorita的转化频率分别可达42.6%和36.8%。 相似文献
77.
78.
Wan-gang GU Jun-fa YUAN Ge-lin XU Li-juan LI Ni LIU Cong ZHANG Jian-hong ZHANG Zheng-li SHI 《Virologica Sinica》2007,22(1):21-25
BALB/c mice were immunized with purified White spot syndrome virus (WSSV). Six monoclonal antibody cell lines were selected by ELISA with VP28 protein expressed in E. coli. in vitro neutralization experiments showed that 4 of them could inhibit the virus infection in crayfish. Western-blot suggested that all these monoclonal antibodies were against the conformational structure of VP28. The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labeling. These monoclonal antibodies could be used to develop immun-ological diagnosis methods for WSSV infection. 相似文献
79.
Yin-ping LU Bao-ju WANG Ji-hua DONG Zhao LIU Shi-he GUAN Meng-ji LU Dong-liang YANG 《Virologica Sinica》2007,22(3):193-198
Guanylate binding protein-1(GBP-1)is an interferon-induced protein.To observe its antiviral effect against Hepatitis B virus(HBV)and Coxsackie virus B3(CVB3),we constructed an eukaryotic expression vector of human GBP-1(hGBP-1).Full-length encoding sequence of hGBP-1 was amplified by long chain RT-PCR and inserted into a pCR2.1 vector,then subcloned into a pCDNA3.1(-)vector.Recombinant hGBP-1 plasmids and pHBV1.3 carrying 1.3-fold genome of HBV were contransfected into HepG2 cells,and inhibition effect of hGBP-1 against HBV replication was observed.Hela cells transfected with recombinant hGBP-1 plasmids were challenged with CVB3,and viral yield in cultures were detected.The results indicated that recombinant eukaryotic expression plasmid of hGBP-1 was constructed successfully and the hGBP-1 gene carried in this plasmid could be efficiently expressed in HepG2 cells and Hela cells.hGBP-1 inhibit CVB3 but not HBV replication in vitro.These results demonstrate that hGBP-1 mediates an antiviral effect against CVB3 but not HBV and perhaps plays an important role in the interferon-mediated antiviral response against CVB3. 相似文献
80.
本文发展的技术可以使金鱼草下胚轴外植体不定芽高频再生。添加有2mg/lBAP和0.5mg/lNAA的MS培养基上再生芽出得又快又多,淡黄色愈伤组织易于形成并存活。但主要依赖的是6-BAP。MS培养基中含有0.1mg/l或6mg/l浓度的6-BAP均能刺激芽的发育,最适浓度为2-3mg/l.仅加2,4-D,也能形成正常的黄色愈伤组织。但这样形成的愈伤组织在只含有0.1-6mg/l6-BAP或含有2mg/l6-BAP和0.5mg/lNAA的MS培养基上均不能分化生出再生芽来。仅加NAA,能够刺激下胚轴形成根。加入AgNO_3,则抑制不定芽的再生,并导致整个外植体变成棕色而死亡。组织化学分析结果表明:不定芽的再生源于下胚轴外植体表皮细胞。在不含有任何生长调节剂的MS培养基上,再生苗均能形成根。再生植株移栽至花盆可开花结实。 相似文献