共查询到20条相似文献，搜索用时 78 毫秒
1.
锥栗自然居群ISSR-PCR分析技术的建立 总被引：3，自引：0，他引：3
提取野生锥栗的基因组DNA作为ISSR-PCR模板,对反应体系中的模板用量、Mg2+浓度、dNTPs浓度、引物浓度和TaqDNA聚合酶用量5个主要影响因素进行梯度试验,并对扩增程序中的退火温度与循环次数进行了探讨,初步确立了适合野生锥栗的ISSR-PCR分析技术体系,即25μL反应体系中,模板DNA40或50ng、Mg2+2.25mmol.L-1、dNTPs0.2mmol.L-1、引物0.2μmol.L-1、TaqDNA聚合酶1.0U;PCR扩增程序为:94℃预变性3min;然后94℃变性45s,(Tm+2～4℃)退火45s,72℃延伸2min,共32个循环;最后72℃充分延伸5min;1.5%琼脂糖凝胶电泳分离扩增产物。 相似文献
2.
以野蔷薇(Rosa multiflora)的冬末初春芽和夏初叶片为材料,研究了DNA的提取方法,并对影响随机扩增多态DNA(RAPD)的反应因素进行了优化.建立了适合野蔷薇RAPD的反应体系和反应程序,在20μL反应体系中,25ng模板DNA,2.0mmol/LMgCl2,0.2mmol/LdNTPs,1UTaqDNA聚合酶,0.1μmol/L引物.扩增程序为:94℃预变性4min,然后94℃变性45s,37℃退火1min,72℃延伸1min,37个循环,最后72℃延伸5min,4℃保存. 相似文献
3.
以内蒙古开鲁县12个红干椒品种为材料,采用改良的CTAB法提取基因组DNA,通过采用正交优化设计方法,对红干椒RAPD反应条件进行了优化.试验结果表明,最佳红干椒RAPD的反应体系(25 μL)为:PCR染料2.0μL,MgCl2 2.0 mmol·L-1,dNTPs 0.6 mmol·L-1,引物0.5μmlo·L-1,DNA模125 ng,Taq酶1.25 U,超纯水14.9μL;适宜扩增条件为:94℃预变性4 min,40个PCR循环94℃变性1 min,37℃退火1 min,72℃延伸1.5 min,72℃复延伸5 min,4℃保存. 相似文献
4.
正交设计对‘红阳’猕猴桃ISSR反应体系的优化研究 总被引：1，自引：0，他引：1
以‘红阳’猕猴桃及其杂交F1代为试材,通过正交设计对ISSR-PCR扩增反应的影响因子进行优化,初步确立了适合‘红阳’猕猴桃ISSR-PCR反应体系:总体积20μL,10×PCR buffer2.0μL,Mg2+0.75 mmol/L,Taq酶1.5 U,模板DNA 60 ng,引物1.25μmol/L,dNTPs 0.15mmol/L。扩增程序:94℃预变性7 min,94℃变性30 s,50~58℃退火45 s,72℃延伸2 min,45个循环,72℃延伸7 min。引物UBC841的最佳退火温度为58.5℃。应用该优化反应体系,用UBC835对35份杂交F1代DNA进行ISSR-PCR扩增,结果显示优化后的反应体系具有较高的稳定性。 相似文献
5.
内蒙古红干椒RAPD反应体系的正交优化 总被引：2，自引：0，他引：2
以内蒙古开鲁县12个红干椒品种为材料,采用改良的CTAB法提取基因组DNA,通过采用正交优化设计方法,对红干椒RAPD反应条件进行了优化。试验结果表明,最佳红干椒RAPD的反应体系(25μL)为:PCR染料2.0μL,MgCl22.0 mmol·L-1,dNTPs0.6 mmol·L-1,引物0.5μmol·L-1,DNA模板125ng,Taq酶1.25U,超纯水14.9μL;适宜扩增条件为:94℃预变性4min,40个PCR循环94℃变性1min,37℃退火1min,72℃延伸1.5min,72℃复延伸5min,4℃保存。 相似文献
6.
菊花SSR-PCR反应体系的建立和优化 总被引：1，自引：0，他引：1
为快速确定菊花SSR反应体系,利用正交实验设计L16(45)对菊花基因组SSR-PCR反应体系的5个因素(模板DNA、Mg2+、dNTP、引物和Taq酶)在4个水平上进行正交设计,筛选出适合菊花的最佳SSR-PCR反应体系,进一步利用单因素完全随机试验筛选各反应因素的最佳水平。结果表明:建立菊花基因组DNA SSR-PCR反应体系为25μL:60ng模板DNA、2.0mmol/L Mg2+、0.1mmol/L dNTP、0.3μmol/L引物、1UTaq酶。并对菊花引物进行梯度退火试验,其最佳退火温度在53.1℃;扩增程序是:95℃预变性5min;32个循环的94℃变性50s、53.1℃退火50s、72℃延伸50s;72℃延伸8min,4℃保存。该体系的建立为今后菊花SSR分析奠定了基础。 相似文献
7.
以辣椒DNA为模板,对影响辣椒RAPD扩增的重要参数进行了优化试验,以期建立辣椒RAPD反应的最佳体系.通过采用正交试验设计的方法,对辣椒RAPD条件进行了优化,结果表明:最佳的辣椒RAPD的反应体系(15μL)中含有1×buffer,MgCl2 2.67 mM,dNTPs 0.13mM,Primer 0.5μmol/L,Taq 0.75 U/μL,DNA 40～50 ng.适宜扩增条件为94℃预变性4 min,再进入40个PCR循环94℃变性1 min,37℃退火45 s,72℃延伸1 min,72℃延伸10 min,4℃保存. 相似文献
8.
采用改良CTAB法提取了桦褐孔菌总DNA,确定ISSR最适25 μL反应体系为模板DNA浓度15 ng·μL-1,dNTPs 浓度150 μmol· L-1,引物浓度25 μmol·L-1,Taq DNA聚合酶浓度2.0 U,Mg2+浓度1.4 mmol· L-1,10×buffer 2.5 μL,其余用ddH2O补足;确定ISSR扩增程序为:94℃预变性5 min,35个循环:94℃变性1 min、45℃～51℃退火1 min(退火温度因不同引物而定)、72℃延伸1 min,最后72℃延伸5 min,4℃保存.筛选出16条ISSR引物,并成功应用引物UBC842完成了21株桦褐孔菌的ISSR-PCR反应. 相似文献
9.
快速微量提取番茄DNA及SSR-PCR反应体系的优化 总被引：1，自引：0，他引：1
以番茄叶片为试材,采用改进的CTAB法,电钻研磨,提取过程中加入醋酸铜,设计4因素3水平的正交试验对PCR反应体系进行优化,同时利用梯度PCR对退火温度进行选择选择。结果表明:优化的10μL反应体系含:1×buffer,1.2 mmol/L Mg2+,1.5 mmol/L dNTPs,1.2μmol/L引物,0.75 UTaqDNA聚合酶,10 ng模板DNA。扩增程序为:94℃预变性2 min;94℃变性30 s,49.2℃退火30 s,68℃延伸30 s,共35个循环;最后72℃延伸8 min。优化的反应体系可以用于SSR分子标记机的研究,在所有SSR引物中基本都能有效扩增;改进的CTAB法提取的DNA纯度更高。 相似文献
10.
通过对TaqDNA聚合酶、Mg2+、dNTP、通用引物、模板DNA浓度等反应参数的系统研究,建立了中国樱桃S-RNase基因特异PCR扩增体系。该体系反应的总体积25μL,其中Taq酶1.25U,MgCl22.5mmol/L,dNTP0.15mmol/L,通用引物0.25μmol/L,模板DNA 50 ng。反应程序为:94℃预变性3 min;33个循环的94℃变性1 min,56℃退火45 s,72℃延伸1.5 min;最后72℃延伸10 min。利用优化的扩增体系在中国樱桃的不同品种中获得有效扩增,进一步证明了该体系具有良好的稳定性和重复性。 相似文献
11.
AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β-particles emission from 188Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by 188Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [3H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of 188Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from 188 Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G0/G1 arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation. 相似文献
12.
AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration. 相似文献
13.
《International Journal of Fruit Science》2013,13(3):101-120
Abstract Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared. 相似文献
14.
The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species. 相似文献
15.
多效唑对猕猴桃离体试管苗生长及内源激素的影响 总被引：18，自引：0，他引：18
多效唑（ＰＰ３３３）处理猕猴桃试管苗，降低了其生长强度；植株体内的ＧＡ３、ＩＡＡ和ＺＴ含量下降，ＡＢＡ的含量上升，乙烯释放率增加；并且能降低外源的ＧＡ３和ＩＡＡ促进生长的作用，而外源的ＧＡ３和ＩＡＡ又能不同程度地逆转多效唑的抑制作用，使植株恢复生长。 相似文献
16.
HU Zhi-cheng ZHU Jia-yuan ZHU Bin GUO Dong CHEN Bin ZHANG Kai HU Kun-hua LI Ming-tao TANG Bing 《园艺学报》2011,27(9):1802-1806
AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars. 相似文献
17.
AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [3-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10-6-10-4 mmol/L) stimulated [3-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [3-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10-6-10-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl2 for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation. 相似文献
18.
Duncan Brean W. Boyle Shannon Breininger David R. Schmalzer Paul A. 《Landscape Ecology》1999,14(3):291-309
Historic landcover dynamics in a scrubby flatwoods (Tel-4) and scrub landscape (Happy Creek) on John F. Kennedy Space Center were measured using aerial images from 1943, 1951, 1958, 1969, 1979, and 1989. Landcover categories were mapped, digitized, geometrically registered, and overlaid in ARC/INFO. Both study sites have been influenced by various land use histories, including periods of range management, fire suppression, and fire management. Several analyses were performed to help understand the effects of past land management on the amount and spatial distribution of landcover within the study sites. A chi-squared analysis showed a significant difference between the frequency of landcover occurrence and management period. Markov chain models were used to project observed changes over a 100-year period; these showed current management practices being effective at Tel-4 (restoring historic landscape structure) and much less effective at Happy Creek. Documenting impacts of past management regimes on landcover has provided important insight into current landscape composition and will provide the basis for improving land management on Kennedy Space Center and elsewhere. 相似文献
19.
AIM: Previous studies performed with XBP-01 in vitro indicated that XBP-01 could inhibit vascular smooth muscle cells from being transformed into foam cell and could eliminate the atherosclerotic plaque in C57BL/6J mouse. This experiment is to investigate its mechanism of eliminating plaques in vitro. METHODS: The cultured porcine artery smooth muscle cells incubated with XBP-01 of 0.1 mg/L for 24 h after preincubated with oxidized low density lipoprotein of 15 mg/L for 72 h in vitro. The samples were analyzed by fluorescence microscope, confocal microscope system and flow cytometry. RESULTS: Apoptosis was triggered by being incubated with oxidized low density lipoprotein and this process was accelerated additionally by being incubated with XBP-01. CONCLUSION: XBP-01 can be effective in eliminating atherosclerotic plaque by accelerating the process in which oxidized low density lipoprotein induced smooth muscle cell apoptosis. 相似文献
20.
XUE Ying-sheng WANG Jing-feng NIE Ru-qiong WU Wei-kang LUO Han-chuan CHEN Xiao-wei 《园艺学报》2004,20(4):603-607
AIM: To investigate the influence of Sini decoction (SND) on the proliferation and apoptosis of rabbit abdominal aorta smooth muscle cells after ballon injury and discuss the effect of vascular smooth muscle cell's (VSMCs) proliferation and apoptosis in post-percutaneous coronary intervention (PCI) restenosis (RS) and the feasibility of SND preventing post-PCI RS. METHODS: The animal model of rabbit abdominal aorta ballon injury was set up and the therapertic group was treated with SND. The shape of proliferative and apoptotic cell were investigated by electron microscope. Immunohistochemistry staining was performed using α-actin,PCNA and Cyclin E monoclonal antibodies. In situ Cell Death Detection Kit was used to identify apoptotic cells. Abdomial aorta angiography was operated in the 84th day subgroup and the stenosis degree was evalued by quantitative angiographic analysis. RESULTS: As compared with the control group, the therapeutic group displayed a lower proliferative percentage and a higher apoptosic percentage (P<0.05). Moreover, the apoptosic peek time was on the 14th day after operation,which was longer than the control group. CONCLUSION: SND effectly inhibited the proliferation of VSMCs and iuduced apoptosis in VSMCs. 相似文献