一种适合芽孢杆菌质粒DNA提取的改良碱裂解法

以Birnboim和Doly的质粒提取方法为基础,通过缩短菌体培养时间,增加菌体收集量,并对提取过程中所用的三种主要溶液的用量进行适当调整等,摸索出质粒提取的一种改良碱法.用改良后的碱法对浸麻芽孢杆菌、地衣芽孢杆菌和枯草芽孢杆菌进行质粒提取,均获成功.     

糖尿病肾病大鼠胰基因组DNA甲基化状态的变化

分别从正常大鼠、糖尿病肾病大鼠、中药糖微康治疗的糖尿病肾病大鼠 、西药开博通治疗的糖尿病肾病大鼠的胰组织中提取其基因组DNA,用对DNA甲基化作用敏感 的限制性核酸内切酶HpaⅡ和MspⅠ(为一组同裂酶)进行限制性酶切;并进行染色质DNaseⅠ敏感性分析. 实验发现中药糖微康治疗后能向正常方向逆转胰组织DNA中CCGG位点甲基化状态,胰组织中处于转录活性状态的染色质明显增加. 这表明中药糖微康能通过提高基因转录水平和向正常方向恢复甲基化状态而发挥其治疗作用.     

DNA计算及DNA计算机的研究进展

概述了DNA计算的基本原理、DNA计算的应用和DNA计算机的研究进展及存在问题，基于DNA生化反应的计算机称为DNA计算机，由于其采用一种完全不同于传统计算机的运算逻辑与存贮方式，DNA计算机在解决某些复杂问题时具有传统计算机无法比拟的优势，目前国际上关于DNA计算和DNA分子生物计算机的研究方兴未艾，极大地推进了DNA计算机的研究过程。     

非损伤性取样的蚕类DNA提取及检测

对家蚕和蓖麻蚕的蚕蜕和蛾翅分别采用SDS裂解法和动物组织基因组试剂盒提取法提取DNS，并与常规方法以蚕蛹提取的DNA一起进行RAPD扩增图谱比较、分析，结果表明用非损伤性方法提取的DNA的RAPD图谱和常规方法提取的DNA的RAPD图谱基本一致。     

阿的平荧光猝灭法测定DNA

在室温条件下，阿的平能嵌入DNA的碱基对之间，而阿的平荧光大幅猝灭，酸性介质下，激发波长348，428或449nm，碱性介质下激发波长为362或422nm,发射波长均为502nm，基于此对核酸进行定量测定，方法简单，快速，灵敏，对于双链或单链小牛胸腺DNA，线性范围0－4μg/mL，检测限0．03μg/mL，共存物质干扰小，在不同浓度ctDNA存在下，温度对荧光光猝灭的影响结果显示猝灭方式为静态猝灭。     

遗传物质稳定性的辩证理解--谈DNA的稳定性与可变性

DNA在生物界中,堪称生物大分子中最重要者,主宰一切生物体维持其生命的各种机能的正常运行及每个物种一代一代繁衍于世,其结构是相对稳定的.     

利用椭圆偏振光谱法研究硅片上DNA分子杂交

将单晶硅片硅烷化后，用戊二醛做醛基修饰，将5′端氨基修饰寡核苷酸探针片段通过醛－氨共价键结合在硅片上，然后用椭圆偏振光谱法对探针和不同序列样品的杂交反应进行了研究。结果表明该方法能在不对样品进行荧光标记的情况下直接研究杂交反应的效率，从而得知样品的序列信息。     

Construction of multivalent DNA vaccines forMycobacterium tuberculosis and its immunogenicity

The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eukaryotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further analysis. Plasmids containing the right insertion were sequenced to confirm their identity. COS7 cells were transfected with a mixture containing serially diluted plasmid DNA encoding three secreted proteins and Lipofectin (Gibco). The supernatants and pellets prepared from various cell lines were run on SDS-PAGE gel and the expression of these proteins in COS7 cells were demonstrated by immunoblot using polyclonal or monoclonal antiserum of M.TBH37Rv. 21 days after first vaccination of C57BL-6 mice by all three recombinant eukaryotic expressing vectors, antibody titer for Ag85B reached 1∶3200. 21 days after second vaccination, the antibody titer reached 1∶102400. The highest antibody levels induced by multivalent vaccines after the second injection were equal to or even greater than the highest antibody levels of single DNA vaccine reported in literature after third injections. Antibody titer of MPT-64 was 1∶50 after the first injection and it reached 1∶200 after the second injection. No antigen-specific antibody against ESAT-6 was detected in sera harvested from immunized mice 21 days after both injections. Antigen-specific IFN-g level of Ag85B was 110 pg/mL while no antigen-specific IFN- g level of ESAT-6 and MPT-64 was detected even after third injections. To our knowledge, it is the first time that studies of polyvalent recombinant DNA vaccines against TB were carried out in C57BL-6 mice. Our results indicated that multiple DNA vaccines could be used to enhance protective responses against M.TB.     

DNA Coding Based Knowledge Discovery Algorithm

A novel DNA coding based knowledge discovery algorithm was proposed, an example which verified its validity was given. It is proved that this algorithm can discover new simplified rules from the original rule set efficiently.     

对于基因芯片实验的动力学范围和结果可重复性的研究

杂交动力学和实验结果的可重复性是基因芯片研究中十分重要的内容，以单基因模拟芯片对DNA芯片的杂交动力学进行了初步的研究：将不同含量的植物cDNA点在玻片上制成DNA芯片，利用体外转录的方法得到植物基因的mRNA，然后通过反转录方法把荧光标记到cDNA上，用不同含量的荧光标记cDNA对芯片进行杂交，根据结果分析了靶基因和探针含量变化对芯片杂交信号产生的影响，利用多基因模拟芯片分析了不同基因的杂交动力学特点；利用12800点的高密度芯片进行了2组不同的表达谱实验，对芯片结果的可重复性做了验证。