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11.
从甜瓜子叶中提取总DNA   总被引:4,自引:1,他引:4  
陆璐  赵长增  陶兴林 《果树学报》2005,22(6):748-750
用分子标记方法检测甜瓜杂交种子纯度,如果从真叶中提取DNA,检测周期至少需要15d。为了缩短检测周期,我们分别以甜瓜的成熟干种子、吸胀水后“露白”种子、3d龄刚转绿子叶、6d龄子叶、9d龄子叶和12d龄子叶为材料,进行了提取总DNA的研究。结果表明:除了干种子和吸胀水后“露白”种子外,其他的材料都可以提取到DNA,但是DNA的质量因子叶日龄不同而存在很大的差异。不同日龄的子叶中,以3d龄子叶提取的DNA质量好。子叶6d龄时,提取的DNA质量次于3d龄,少量DNA开始出现降解,以后随着子叶日龄的增加,DNA降解加重。另外,与以真叶为材料提取的DNA样品相比较,子叶DNA样品中的蛋白质等杂质含量高,应增加氯仿抽提纯化次数。  相似文献   
12.
在不同提取方法下蜜蜂基因组DNA浓度的比较   总被引:7,自引:1,他引:7  
吉挺  陈晶  潘瑞 《中国蜂业》2005,56(12):10-11
本研究采用了国内外常见的5种蜜蜂基因组DNA抽提的方法,对意大利蜂工蜂基因组DNA进行提取,并进行了浓度检测.其中方法1,方法2采用的是5日龄的工蜂蛹体,而其他3种方法采用的是成年工蜂的头胸部进行提取.试验结果表明这5种方法之间差异极显著,其中方法2的浓度最高,为84.7850±24.5974,说明在相同实验条件下,方法2(蜜蜂蛹体一次抽提法)在浓度层次上效果最好.  相似文献   
13.
AIM:To explore the mechanism of neuronal injury and repair by investigating the expression of caspase-3 and apurinic/apyrimidinic endonuclease (APE/Ref-1) after focal cerebral ischemia. METHODS:A model of middle cerebral artery occlusion in rats was performed. The expression of caspase-3P20 and APE/Ref-1 was examined by immunohistochemistry staining, TUNEL was applied to detected DNA damage, and double labeling with TUNEL and APE/Ref-1 was used to determine the relationship between APE/Ref-1 and DNA damage. RESULTS:The active subunit P20 of caspase-3 was predominantly expressed within ischemic penumbra. The peak time of caspase-3P20 positive cells preceded the appearance of TUNEL. With aggravation of cerebral ischemia, APE/Ref-1 immunoreactive cells in penumbra were significantly decreased. CONCLUSION:The activation of caspase enzymatic cascade following cerebral ischemia leads to degradation in DNA, meanwhile, decrease in DNA repair molecules or the failure of DNA repair may deteriorate the course.  相似文献   
14.
The major role of DNA polymerase β was thought to be limited in its involvement in short patch base excision repair by removing 5’-deoxyribose phosphate and base insertion. However, the recent researches indicate that polymerase β might take part in a wide spectrum of DNA metabolism reactions, including long patch base excision repair, DNA replication, recombination, meiosis and transleisional DNA synthesis. Because of its wide and important cellular function, an inappropriate intracellular polymerase β level might be associated with genomic instability. Down-regulation or mutation of polymerase β is mutagenic due to deficient in DNA repair, while overexpression of this error-prone β polymerase might perturb the normal function of other accurate polymerases and cause genomic instability as well.  相似文献   
15.
AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [3H]-TdR incorporation and [3H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts.  相似文献   
16.
昆虫病原线虫rDNA多态性分析   总被引:1,自引:0,他引:1  
本文对国内外昆虫病原线虫斯氏属和异小杆属的47个品系进行rDNA—ITS PCR—RFLP分析,研究其DNA多态性,并构建了分子系统发育树状图。各品系的ITS区无明显的长度差异,PCR—RFLP分析将47个品系分为斯氏属和异小杆属两大类,两属线虫又分别分为11组和4组。所得结果丰富了ITS PCR—RFLP图谱库,为弄清我国的昆虫病原线虫与国外种类的分子系统发育关系及未定名线虫的鉴定提供重要依据,同时为筛选适合的线虫种类防治害虫奠定基础。  相似文献   
17.
The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.  相似文献   
18.
利用室内选育119代的抗溴氰菊酯小菜蛾Plutella xylostella品系和敏感品系,通过反复回交和自交建立近等基因系。用Operon公司合成的20个引物对近等基因系基因组DNA进行PCR扩增。3个引物在抗性品系中分别产生1条特异扩增带,4个引物在敏感品系中分别产生了1~5条特异扩增带。由于在近等基因系中,除了与抗性相关的基因区外,其它的遗传背景是相同的,因此可以认为这些特异带与小菜蛾对溴氰菊酯的抗性有关。  相似文献   
19.
Clones of Hylocereus and of Selenicereus species were distinguished from each other by the banding pattern generated by one to nine 10-mer oligonucleotide primers in the random amplified polymorphic DNA (RAPD) reaction. RAPD analysis was also applied to estimate the genetic relationship among five Hylocereus and nine Selenicereus species. A dendrogram was constructed based on a data matrix of 173 polymorphic bands originated by nine primers. Two groups were identified, one consisting of Hylocereus species and the other consisting of Selenicereus species. These results are consistent with the accepted taxonomic classification of the genera studied. The principal coordinate analysis (PCO), i.e. the plot drawn on the basis of the RAPD data, clearly distinguished between three groups, namely, Hylocereus species, S. megalanthus and the rest of the Selenicereus species studied. PCO thus strongly support the notion that the tetraploid S. megalanthus is an exception among the Selenicereus group. The RAPD results support our hypothesis regarding the allopolyploid (rather than autopolyploid) origin of this species.  相似文献   
20.
AIM: To investigate the expression of nucleotide excision repair gene ERCC1 and its relationship with PAH (polycyclic aromatic hydrocarbons)-DNA adducts in lung cancer tissues. METHODS: ERCC1 mRNA expression and the PAH-induced DNA adducts were detected in 150 lung cancer tissues, 120 adjacent lung tissues without cancer cells, 40 benign lung lesions and 40 normal lung tissues. The effects of some exposure factors on the expression of ERCC1 gene and the connection between ERCC1 and PAH-DNA adduct was analyzed. RESULTS: Reduced expression levels of ERCC1 were observed in 46 of 150 (30.7%) lung cancer specimens and 1 of 40 (2.5%) normal lung tissues. Smoking may suppress the expression of ERCC1 gene. The level of PAH-DNA adduct was negatively correlated with the expression of ERCC1 gene, the Spearman coefficient was -0.648, P<0.01. CONCLUSION: ERCC1 is an important nucleotide excision repair gene and may participate in the repair of DNA damage, such as PAH-DNA adduct. Low expression of ERCC1 may play an important role in the development of human lung cancer.  相似文献   
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