中国部分地区绵羊肺炎支原体的基因多态性分析

为了解绵羊肺炎支原体(Mycoplasma ovi pneumoniae,Mo)在我国部分地区的流行病学特征,采用随机扩增多态性DNA(RAPD)及限制性片段长度多态性(RFLP)方法对26株Mo基因多态性进行了研究,并利用NTsys2.10e软件对获得的多态性图谱进行了聚类分析.结果在相似系数为0.70时,26株Mo可分成6个RAPD群或6个RFLP群;相似系数为0.90时,分成18个RAPD群或18个RFLP群;相似系数为1.00时,分成25个RAPD群或26个RFLP群.结果表明,我国Mo存在高度的基因多态性,这种多态性与地域差异相关,与宿主来源也具有一定的相关性.研究结果为了解我国Mo的分子流行病学特征打下了基础,也为建立有效的Mo诊断方法和疫苗研制提供了参考依据.     

PCR-ELISA法对大豆品种的转基因定性检测研究

以共价交联在PCR管壁上的寡核苷酸作为固相引物进行PCR扩增,对PCR扩增的固相和液相产物分别进行杂交和凝胶电泳检测的PCR-ELISA法,对黑龙江省常规选育品种合丰35和黑农37、外源DNA直接导入大豆的分子育种所获品种黑生101、及大连进口的美国2号等大豆,进行转基因定性检测.结果表明:该方法高效可靠,可作为一种快速定性检测转基因产品的方法;检测结果:美国2号为转基因大豆,黑生101、合丰35和黑农37为非转基因大豆.上述结果对分子育种、生态保护、安全监测及对建立黑龙江省非转基因大豆生产保护区等具有重要意义.     

Analysis of DNA Methylation and mRNA Expression Level of MSTN Gene in Xinjiang Bashiby Sheep

In order to investigate DNA methylation and expression levels of myostatin (MSTN) gene in mscule and fat, 5 months of age of Bashiby sheep were selected, the promoter region and exon 1 methylation levels of MSTN gene was analyzed using bisulfite sequencing PCR (BSP). Real-time PCR was used to detect the expression level of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat of Bashiby sheep. The results showed that the methylation probability of muscle was higher than fat, the methylation probability of biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle and tail fat were 74.2%, 74.2%, 83.2%, 83.7%, 82.1% and 25.3%, respectively. The expression levels of MSTN gene in biceps femoris, femoral triceps, semitendinosus, semimembranosus, longissimus dorsi muscle were significantly lower than tail fat in Bashiby sheep (P < 0.05), but there were no significant difference among biceps femoris, femoral triceps, semitendinosus, semimembranosus and longissimus dorsi muscle (P > 0.05).The DNA methylation was negatively correlated with the expression levels in muscle and fat of Bashiby sheep (r=-0.886, P < 0.05).     

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.     

Detecting<Emphasis Type="Italic">Orobanche</Emphasis> species by using cpDNA diagnostic markers

Some species of the genusOrobanche are among the most devastating parasitic weeds, causing extensive damage in agricultural fields. Considering the difficult control due to seed longevity in the soil, small seed size, high fecundity and a subterranean phase that allows them to parasitize the host before they emerge and become evident, the development of diagnostic markers is highly recommended. In our study we identified potential molecular diagnostic markers from the plastid genome in order to distinguish among the most importantOrobanche species attacking crops in Andalusia, the southern region of the Iberian Peninsula. The study has consideredO. crenata, O ramosa andO. cumana causing serious losses in legumes, solanaceous crops and sunflower fields, respectively, andO. minor that, although abundant in Andalusia, has to our knowledge not yet been found parasitizing agricultural hosts. We amplified a non-coding region from the plastid genome, studied sequence differences among the amplified fragments and digested those of the same length with selected restriction enzymes. Here, we propose a molecular protocol to distinguish the main parasitic plants in crop fields of southern Spain. Different applications such as identification ofOrobanche seeds in soil or crop seed lots are discussed in order to offer right crop recommendations or to prevent new infestation of parasite-free fields. Recommendations for further development of these diagnostic markers are also considered. http://www.phytoparasitica.org posting Jan. 15, 2007.     

DNA指纹技术对近交系小鼠生产扩大群的遗传检测

采用生物素标记的（GGAT）4寡核苷酸探针对C57BL／6J、BALB／c和DBA／2三种近交系生产扩大群F4代小鼠进行了DNA指纹图分析。结果显示（GGAT）4寡核苷酸探针对上述三种近交系小鼠产生的DNA指纹图的图带数均为10-12条,具有良好的多态性,品系内平均DNA指纹图的相似系教（X）在0．88—0．95的范围内,具有相同指纹图的概率（P）均在0．35以上,极显著地高于品系间的相似系数（0．18—0．31）和相同指纹图的概率（P〈8．4&#215;10^-7）。研究结果表明（GGAT）。寡核苷酸探针可用于制作近交系小鼠生产扩大群的DNA指纹图,以对其进行遗传检测。     

耦合水文情势及鱼类繁殖的江垭水库生态调度需求研究（生态调度专刊）

大型水库实施生态调度是修复河流水文情势、维持河流生态系统健康的重要手段。江垭水库位于溇水干流下游的张家界慈利县，是澧水流域具有重要防洪意义的大型水库。本研究通过环境DNA采样、IHA水文指标变动分析和文献资料总结等方法，开展了江垭坝下水文情势及鱼类繁殖需求的关系研究，提出了江垭水库生态调度需求及建议。2021年9月在溇水干支流综合调查到鱼类55种，与上世纪90年代相比，流水性种类在干流江段显著减少了51.35%；按照生态调度优先等级评估原则，确定江垭水库生态调度的主要目标物种为产漂流性卵的银鲴、银鮈、贝氏?和产粘沉性卵的鲤、鲫；江垭建坝前后下游水文情势变化较大，发生高度改变且对鱼类洄游产卵及鱼卵发育有重要作用的水文指标包括：年最大1日、3日平均流量指标，年出现高流量脉冲事件的次数和持续时间，以及连续日流量上涨率。以促进坝下不同产卵类型的鱼类繁殖为目标，江垭水库生态调度一方面需要恢复一定的高流量脉冲次数和历时，另一方面需要控制下游的水位日降幅，通过泄放合理的生态流量来维持下游鱼类的生物多样性。     

RAPD技术在产气荚膜杆菌基因分型中的应用研究

采用随机扩增多态性ＤＮＡ技术对产气荚膜杆菌进行ＰＣＲ扩增。结果产气荚膜杆菌Ａ，Ｂ，Ｃ，Ｄ４个型均可扩增出约４３０ｂｐ的条带，Ｂ，Ｃ型还扩增出约２１０ｂｐ条带，Ｂ，Ｄ型还扩增出约１２１０ｂｐ条带，Ａ型还扩增出约２４０ｂｐ和６９０ｂｐ条带，各型之间的条带差别明显，从而为产气荚膜杆菌的基因分型、ＰＣＲ快速诊断提供可靠的依据。     

Polymorphism in Mitochondrial DNA of Six Fowl Breeds Revealed by Restriction Endonuclease

The patterns of cleavage of mtDNA by restriction endonucleases was analysed for six fowl breeds and of .Hyline White,Hyline Brown,ISA Brown,Hisex Brown,Lohmann White,Abor Acres and mtDNA polymorphisms were detected in the restriction patterns with the following eight enzymes,Ava Ⅰ,AvaⅡ,EcoRⅠ,HindⅢ,BamHI,PvuⅡ,PstⅠ,HincⅡ.The restriction cieavage patterns were identical among these breeds.Hyline White,Hyline Brown,ISA Brown,Hisex Brown,Lohmann White-A type.The patterns of Abor Acres were Btype.Based on their mtDNA restriction types,all the breeds were classified into two groups.Genetic distances among these groups were calculated in roder to define their phylogenetic relationships.The relationship among five egg line breeds is close,while Abor Acres(Broiler fow)is relatively far from them.The results suggest that the difference of mtDNA could result from the different origins.The polymorphic sites in mtDNA of Hyline White bas been located on a restriction map.     

3种提取基因组DNA方法的比较分析

分别采用3种不同的方法提取基因组DNA，比较所提取的DNA的质量以及提取所需时间、费用，以期选择一种高效、简洁快速、价格合理的提取方法。结果发现这3种方法提取的质量都能达到相关分子生物学试验的要求，但在操作步骤、时间、价格等方面存在差异，具体比较结果可为分子生物学试验选择提取方法提供依据。