共查询到20条相似文献，搜索用时 449 毫秒
1.
抑癌基因PTEN载体构建及在LOVO细胞中的表达 总被引：1，自引：0，他引：1
目的:构建抑癌基因PTEN的表达载体并在LOVO细胞中表达。方法:从人外周血中提取总RNA,通过RT-PCR扩增出PTEN基因片段,将PTEN基因连入pLenti6/V5载体。测序鉴定后,经脂质体转染入LOVO细胞,对细胞株进行RT-PCR和Western blot检测。结果:DNA测序证实pLenti6/V5-PTEN表达载体为阳性克隆;该表达载体转染LOVO细胞后上调了PTEN mRNA和蛋白的表达。结论:成功构建了pLen-ti6/V5-PTEN表达载体,为进一步研究PTEN在LOVO细胞中的功能奠定了基础。 相似文献
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用电泳纯的小麦多聚半乳糖醛酸酶抑制蛋白免疫新西兰雄兔,获得了特异性强,效价较高的小麦.多聚半乳糖醛酸酶抑制蛋白抗血清.用此抗血清采用Dot—immunobinding assay,对流免疫电泳二种方法测定PGIP的检出限量,由此建立了多聚半糖醛酸酶抑制蛋白的Dot—immunobinding检测方法,并对制备该抗血清的意义予以讨论. 相似文献
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从人直肠癌细胞株Colo320中提取总RNA,经RT-PCR扩增出SOD基因片段,经限制性内切酶（BamHI,Sinai）酶切后按正确的读码框顺序插入到pGEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCR和质粒双酶切鉴定、序列测定确认,证实成功地构建了人SOD基因融合表达载体．转化菌经IPTG诱导表达,SDS-PAGE显示有与预期大小约44kD相吻合的融合蛋白带,获得了可溶性表达的目的蛋白,采用Glutathione Sepha—rose4B亲和层析对重组蛋白进行纯化,获得了纯度很高的目的蛋白． 相似文献
4.
从水生植物凤眼莲叶片中提取总 RNA,经 RT-PCR扩增出Ca2＋-ATPase基因片段,经限制性内切酶(Sma I,Not I)酶切后按正确的读码框顺序插入到pGEX-4T-2表达载体上,重组质粒转化大肠杆菌,经菌落PCR和质粒双酶切鉴定、序列测定确认,证实成功地构建了Ca2＋-ATPase基因融合表达载体,．转化菌经IPTG诱导表达,获得了大小约48 kD的可溶性目的蛋白,与预期相吻合．利用谷胱甘肽琼脂糖凝胶4B(Gluta-thione Sepharose 4B)亲和介质对重组蛋白进行纯化,获得了高纯度的目的蛋白． 相似文献
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目的：克隆人Lumican基因及构建Lumican基因真核表达载体并研究其对人子宫内膜癌细胞HEC-1A增殖的影响。方法：取人新鲜正常子宫内膜组织,提取总RNA,采用逆转录-聚合酶链反应（RT-PCR）扩增Lumican基因,将该基因定向克隆到真核表达载体pEGFP-N1中,构建真核细胞表达载体pEGFP-N1-Lumican,用限制性内切酶酶切分析,DNA序列分析鉴定重组质粒;将测序正确的Lumican基因通过脂质体介导下转染人子宫内膜癌细胞HEC-1A;逆转录-聚合酶链反应（RT-PCR）和Western Blot 检测转染细胞HEC-1A的Lumican mRNA和蛋白表达。采用MTT法观察转染Lumican基因的人子宫内膜癌细胞HEC-1A的增殖能力。结果：重组质粒pEGFP-N1-Lumican经HindⅢ和BamHⅠ酶切,获得8311 bp片段和865 bp插入片段,序列分析表明插入的片段与Gene Bank发布的序列一致;荧光显微镜下可见转染的HEC-1A细胞有绿色荧光蛋白的表达;转染细胞HEC-1A的Lumican mRNA表达上调,Lumican蛋白相对上调率为74．62％（P＜0．05）。与对照组细胞比较,转染Lumican 基因的人子宫内膜癌细胞HEC-1A的抑制率为21．35％±2．78％。结论：成功构建了真核表达载体pEGFP-N1-Lumican,该载体在体外能有效抑制人子宫内膜癌细胞HEC-1A的增殖能力,为以Lumican基因为靶点研究其对人子宫内膜癌细胞HEC-1A的恶性生物学特性提供实验基础。 相似文献
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饱和苯酚氯仿法提取Tg2576转基因鼠基因组DNA,PCR法扩增编码β-淀粉样蛋白的目的基因,利用基因克隆技术构建以β-淀粉样蛋白为靶的表达载体pcDNA3.1-Aβ42×2,应用酶切及测序鉴定表达载体.相应的双酶酶切能够获得插入的目的基因片段(为269bp),测序未发现突变,表达载体构建成功. 相似文献
7.
目的构建L-ficolin突变体的原核表达载体，为研究L-ficolin的糖结合位点奠定基础。方法设计L-ficolin突变体基因上、下游引物，分别引入EcoRⅠ和XhoⅠ酶切位点。以插入L—ficolin M2（Val144Phe）和M6（Glu 307Asp）点突变的完整编码区基因的质粒pCDNA3-L-ficolin为模板，通过PCR扩增获得突变体的基因片段，将其克隆入原核表达载体pGEX—KG．然后通过酶切和序列测定对其进行鉴定；将此表达载体转化入表达菌BL21（DE3）pLvSs诱导表达。并采用SDS—PAGE对表达产物进行鉴定。结果经酶切及序列分析表明．成功地构建了重组原核表达载体pGEX-KG—L—ficolin—M2、pGEX-KG-L-ficolin—M6，SDS—PAGE显示目的蛋白大量表达。结论L—ficolin突变体融合蛋白可以在大肠杆菌中大量表达。为研究L-ficolin的糖结合位点打下基础。 相似文献
8.
孙晓东 《陕西教育学院学报》2010,26(2):106-107,121
为了构建人C6orf210基因原核表达载体,并在E．coliBL21中表达并纯化。采用RT—PCR扩增人C6orf210基因片段,并将其克隆到原核表达载体Pet21a中,构建重组质粒pET21a—C6ort210。经限制性内切酶EcoRI与XhoI双酶切鉴定及序列测定后,转化E．coliBL21,经IPTG诱导表达融合蛋白。结果显示获得全长为1188bp的人的CAiorf210基因片段。以构建的重组质粒pET21a—C6orf210转化E．coliBL21后,经IPTG诱导,表达出相对分子质量（Mr）约66000的融合蛋白。经SINS—PAGE、Western blot分析显示,诱导表达的蛋白为C6orf210。 相似文献
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《邢台职业技术学院学报》2019,(5):71-74
本研究以不同时期狗头枣果实为材料,测定了纤维素酶(Cx-cellulase)、β-半乳糖苷酶(β-Gal)、果胶甲酯酶(PME)、多聚半乳糖醛酸酶(PG)等4种细胞壁酶的活性。结果表明:Cx-cellulase的活性在枣果实生长发育期间增长趋势平缓,从幼果期到完熟期大约增加1.7倍;β-Gal在枣果实幼果期活性较高,由全红期到完熟期大约增加4.6倍;PME的活性由幼果期到泛红期大约增加1.6倍,泛红期到全红期下降3.5倍,之后趋于平缓;PG的活性从幼果期到成熟期增长较缓慢,成熟期到完熟期活性迅速上升,其活性大约增加1.3倍。在成熟期,PG和Cx-cellulase活性的升高与枣果实软化密切相关,而PME则与枣果实成熟间接相关,β-Gal与枣果实成熟前期软化启动密切相关,在成熟后期促进了果实的软化。 相似文献
11.
Wang AR Lin WW Chen XT Lu GD Zhou J Wang ZH 《Journal of Zhejiang University. Science. B》2008,9(10):818-822
A new stem rot disease is found to occur naturally on Arabidopsis plants in greenhouses of Fuzhou, China. In order to identify its pathogen, we conducted a series of fimgal isolation and purification, plant reinoculation, and ascus and ascospore induction from the sclerotia. The isolate caused typical water-soaked lesions after reinoeulation and produced sclerotia both on Arabidopsis plants and culture medium plates, and the sclerotia could be induced to produce discal apothecia and 8 binucleate ascospores per ascus. These disease symptom and fungal morphology data revealed that the fungus Sclerotinia sclerotiorum (Lib.) de Bary was the pathogen for Arabidopsis stem rot. To confirm this, we further amplified its large subunit ribosomal DNA (LSU rDNA) by polymerase chain reaction (PCR), and compared the sequence with the known LSU rDNA sequences in GenBank. The results show that the sequence shares the highest identities with the LSU rDNAs of different S. sclerotiorum strains. Taking all these data together, we concluded that the fungus that caused the Arabidopsis stem rot is S. sclerotiorum (Lib.) de Bary. This is the first report that Arabidopsis is naturally infected by S. sclerotiorum. 相似文献
12.
短毛独活精油提取及抑菌作用研究 总被引：3，自引：0，他引：3
张知侠 《咸阳师范学院学报》2007,22(2):33-34
用石油醚提取获得短毛独活精油,采用生长速率法测试短毛独活精油的杀菌活性。结果表明:短毛独活精油对12种供试真菌均具有不同程度的抑菌作用,其中对小麦纹枯病菌的抑制率最强,达到100%;其次是油菜菌核病菌、辣椒疫霉病菌,达到80%以上;而对玉米大斑病菌、稻瘟病菌的抑制率相对较低。 相似文献
13.
为了探讨时间复制任务中是否存在编码方式（连续编码和中断编码）和中断位置效应,实验1和2分别考察编码阶段有无中断和中断位置对时间复制成绩的影响。结果发现,对于1700ms靶时距而言,两种编码方式条件下的时间复制成绩无显著差异;对于2300ms靶时距而言,中断编码任务条件下显著较连续编码任务条件下时间复制长度短。而且,不管对1700ms还是2300ms靶时距而言,随着中断位置越靠后,时间复制长度都越短。这说明,编码中断及中断位置对时间复制有显著影响,时间复制任务中存在中断位置效应。 相似文献
14.
Fruit ripening is a complex process and is regulated by many factors.Ethylene and polygalacturonase (PG),lipoxygenase (LOX),expansin (EXP) are all critical regulating factors in fruit ripening and softening process.With antisense,ACS tomato,Nr mutant tomato and cultivated tomato as materials,Northern blot hybridization showed that PG,LeEXP1 and LOX expressed differently in different parts of cultivated tomato fruit during ripening,which was related to fruit ripening.The ripening process of columella and radial pericarp was faster than pericarp.In both Nr mutant and antisense ACS transgenic tomato fruit,expression levels of PG,LeEXP1 and LOX were generally lower than those in cultivated fruit but still related to fruit ripening.The expression levels of PG,LeEXP1 and LOX increased in the mature green tomato fruits after 0.5 h treatment with ethylene ( 100 μL/L).These results indicate that gene expression of PG,LeEXP1 and LOXwere positively regulated by ethylene.The time and cumulative effect of the concentration exists in the expression of PG regulated by ethylene.The regulation of LOX expression mainly depended on the fruit development after great amount of ethylene was produced.PG played a major role in ripening and softening of tomato fruit,and cooperated with the regulation of EXP and LOX. 相似文献
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刘咏莲 《安徽师范大学学报(人文社会科学版)》2003,31(2):245-248
聂耳的歌曲创作有两个重要艺术成就和特色 :一是在群众歌曲这一体裁中 ,善于通过丰富多彩的形象塑造 ,反映生活现实和思想感情 ,特别是准确鲜明地反映 2 0世纪 30年代中国工人阶级和广大劳动群众的形象与感情 ;二是在音乐语言的运用上 ,独具新意和个性 ,既具有民族特色 ,又富于时代气息 ,并将曲调的平易性与独创性有机结合 ,成为广大群众所“喜唱乐闻”的真正的“群众歌曲”。 相似文献
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Michelle Morgan 《高等教育研究与发展》2014,33(6):1150-1165
The global growth in postgraduate (PG) study since the mid-1990s has been attributed to the expansion in Masters by Coursework participation (Bekhradnia, B. (2005). Postgraduate education in the UK: Trends and challenges higher education policy institute. Paper presented at a conference The future of postgraduate education supporting the students of today and tomorrow, London, 17 March). However, unlike at undergraduate level, research into understanding PG growth has been under-researched. This paper aims to contribute to the knowledge gap by identifying and comparing the growth in PG study in Australia, Canada, the USA and the UK. It explores the possible drivers behind the growth and concludes by highlighting potential challenges facing the future of PG study across the sector. 相似文献
17.
三羟甲基乙烷、新戊二醇及其二元体系固—固相变贮热的动力学研究 总被引：1，自引：0，他引：1
运用kissinger及Ozawa方法两种热分析动力学模型,利用差示扫描量热法(DSC)测定三羟基乙烷(PG)、新戊二醇(NPG)及其二元体系的低温固一固相变DSC曲线,求算固一固相变的表观活化能和反应级数,探讨表观活化能随组成不同的变化规律。两种方法的处理结果是相一致的。 相似文献
18.
对本钢连轧厂精轧第七架2*2300kw直流轧钢电机电枢铁心松动故障,进行事故原因分析,找出快速修复的方法。 相似文献
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Dennis A. Vanderweele 《Learning & behavior》1974,2(4):309-312
Mongolian gerbils (Meriones unguiculatus) were examined for components of thirst in the laboratory. The gerbil, reputed to be a desert-adapted rodent, was tested for its response to quinine adulterations of water, hypertonic solutions (1 M sodium chloride, NaCl), and hyperoncotic solutions (40% W/V polyethylene glycol in isotonic saline, PG). Both NaCl and PG were administered intraperitoneally. Gerbils accepted quinine when it was the only solution available and food was ad lib, but preferred distilled water to quinine if both were available. Both PG and NaCl produced increased water intakes in gerbils within the first 2 h, postinjection. PG, however, produced the most reliable and largest increase in water consumption, illustrating that extracellular dehydration is probably a more significant thirst stimulus in gerbils. 相似文献