利用简化的SDS法提取杨树基因组DNA

以杨树组培苗幼嫩叶片为材料,在室温下采用简化的SDS法快速制备其基因组DNA,并对制备DNA过程中的影响因子进行了初步研究。结果表明,采用此方法制备的基因组DNA在数量和纯度上可以满足聚合酶链式反应(PCR)的要求。另外,DNA提取缓冲液中添加一定浓度的PVP、β-巯基乙醇,有利于DNA提取。     

四种桉树青枯菌DNA提取方法及PCR检测灵敏度比较

以带有青枯菌Ralstonia solanacearum的桉树组织为材料,采用4种不同的提取方法抽提青枯菌DNA,同时利用青枯菌的特异性引物进行PCR扩增,比较了4种DNA的提取方法及PCR检测的灵敏度。结果表明,煮沸法和热裂解法操作相对简单,但检测灵敏度较低,检测限分别为104CFU/mL和103CFU/mL;简化提取法和试剂盒法检测效果较好,均可检测到102CFU/mL的青枯菌;简化提取法比试剂盒法操作更简单,检测成本低,具有更高的应用价值。     

一种适于提取复烤烟叶DNA的方法

以改良CTAB法提取复烤烟叶DNA，经U－3000紫外分光光度计测定，所获DNA呈典型的吸收曲线，且OD260／OD280在1．7－1．9之间；对烟草叶绿体不编码序列进行PCR扩增，获得理想条带。     

漾濞核桃叶片基因组DNA的两种提取方法效果比较

以漾濞核桃不同生长期的叶片为材料，采用高盐低pH法和改良CTAB法作提取其基因组DNA的效果比较实验。通过琼脂糖凝胶电泳、紫外分光光度计和RAPD扩增对所提取的DNA样品进行检测，比较所得DNA的产量、质量，认为从漾濞核桃不同发育时期叶片中获得DNA的提取效果不同。4种叶样以冬芽和新叶的效果为好，老叶最差；用两种提取方法从叶片中得到的DNA产量和质量有所不同，高盐低pH法提取的DNA纯度高，但是产量较低；而改良CTAB法则相反，产量高，纯度低。     

松树种子胚乳DNA的抽提与分析

运用改良SDS(十二烷基硫酸钠)法,从马尾松、黄山松、黑松等松树种子胚乳中提取到较高纯度的DNA,并对其纯度、浓度及产率进行了分析.经RAPD检测表明,DNA扩增效果良好,完全能满足常规DNA实验的要求,为针叶树种等其它小粒种子DNA的提取提供了经济、快速、可靠的实验方法.     

Wood identification of JapaneseCyclobalanopsis species (Fagaceae) based on DNA polymorphism of the intergenic spacer betweentrnT andtrnL 5′ exon

DNA was extracted from wood samples of six representativeCyclobalanopsis species (Fagaceae) growing in Japan that cannot be distinguished from one another by conventional microscopy. A part of the intergenic spacer region betweentrnT andtrnT 5 exon was amplified and sequenced. The sequences obtained from wood samples were grouped into three DNA types by a single nucleotide polymorphism as reported previously in leaf samples: I (Quercus acuta, Q. sessilifolia, Q. salicina), II (Q. myrsinaefolia, Q. glauca), and III (Q. gilva). Thus,Q. gilva can be distinguished from the otherQuercus species, and the others are separated in two subgroups based on DNA polymorphism. The present findings support the possibility of wood identification based on DNA polymorphism.     

Association between methylation status of apoptosis-related genes and chemosensitivity of lung adenocarcinoma cell line P15

AIM: To investigate the association between methylation status of apoptosis-related genes and chemosensitivity in the lung adenocarcinoma cell line P15.METHODS: Methylation-specific PCR was applied to detect the methylation status of p73, p14^ARF, p16^INK4a and bax genes of P15 cells in untreated control group and decitabine (DAC) treatment group. RT-PCR was used to detect the expression of p73, bcl-xL, bad, bax, p14^ARF and p16^INK4a at mRNA level. Colony formation assay and cell growth inhibition assay were used to detect the sensitivity of P15 cells to cis-diaminedichloroplatinum (C-DDP) before and after DAC treatment. DAPI staining was used to determine the apoptosis of P15 cells exposed to C-DDP before and after DAC treatment. RESULTS: p73, p16^INK4a and bax were expressed in the methylation status. After DAC treatment, p16^INK4a expression was decreased, and the expression of p73 and bax disappeared. The expression of p73, p16^INK4a and bax in the unmethylated status was weak, but the enhanced expression was observed following DAC treatment. After P15 cells were treated with DAC and C-DDP, the colony formation rate of the P15 cells was significantly decreased as compared with untreated control group. The apoptotic P15 cells in DAC+C-DDP treatment group were significantly higher than those in untreated control group (P<0.05). CONCLUSION: After treated with DAC, the sensitivity of P15 cells to C-DDP is increased due to the activation of silenced pro-apoptotic genes. DAC and C-DDP synergistically promote tumor cell apoptosis. They have significant anti-tumor effect.     

常山胡柚叶片DNA的抽提与分析

经反复实验，采用在抽提前加入一定量的抗氧化剂β－巯基乙醇，用冰冻的异丙醇来沉淀DNA，并增加氯仿／异戊醇抽提次数的所谓改良CTAB法，对常山胡柚叶片DNA抽提与分析，不仅成功提取到常山胡柚叶片DNA，而且所得的DNA浓度高、纯度好，20个试样的DNA苹均质量浓度为272.86μg/mL，平均每克鲜叶片可获DNA约82μg，纯度基本上都在1.4-1.8范围内，并经RAPD反应验证，完全能满足常规分子生物学分析的要求。     

DNA分子标记技术在食用菌研究中的应用及进展

根据近年来国内外的文献报道,对DNA分子标记技术在食用菌遗传育种、菌种和菌株鉴定、遗传多样性分析、基因定位和克隆等研究中的应用及进展进行了综述和分析.认为随着分子生物学技术的进一步发展,将有更多、更有效的分子标记在食用菌研究中得以应用,同时随着分子标记技术应用的深入,更高密度、更为实用有效的遗传连锁图谱将被建立,大量重要的功能基因将被定位和克隆,这将为进一步培育高产、优质、抗病、耐贮等食用菌新品种奠定良好的基础.     

基于DNA条形码技术的泰国番石榴中实蝇幼虫分子鉴定研究

实蝇是世界重要的检疫性害虫之一,对果蔬生产及其国际贸易具有很大的影响。而以线粒体DNA细胞色素C氧化酶亚单位I(mtDNACOI)基因的部分序列作为实蝇的DNA条形码能减少对实蝇成虫形态特征的依赖,可检测其任何虫态的样品,有助于实蝇样品的快速鉴定。本研究采用DNA条形码技术,针对采自泰国四色菊市番石榴烂果中的5头实蝇幼虫进行COI扩增测序,与生命条形码数据库(BOLD)中的序列进行比对并利用PAUP4.0软件构建了其系统进化树。根据序列分析和系统进化关系分析的结果,将5头实蝇幼虫样品鉴定为番石榴实蝇(Bactrocera correctaBezzi),并将本研究获得的2条COI序列在GenBank中注册,GenBank的登录号为HM590450和HM590451。