现代生物工程技术简介

对组成现代生物工程的六大技术系统：基因工程、蛋白质工程、酶工程、细胞工程、微生物工程、生化工程的历史和现状，工作内容及应用前景作了简明的介绍。     

表达HIV-1 Gag-Pol抗原的重组痘病毒的构建及免疫原性

目的构建稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体疫苗,并检测其体液免疫效果。方法将修饰型HIV-1Gag-Pol基因插入到痘病毒表达载体pSC11m2启动子P7·5下游,经与野生型痘病毒同源重组后,进行蓝白斑筛选。用筛选得到的重组病毒rM-GP转染HEK293细胞,经SDS-PAGE和Westernblot检测表达蛋白。用该重组病毒免疫BALB/c小鼠,并检测体液免疫应答。结果已筛选到稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体rM-GP,经Westernblot,在免疫小鼠血清中检测到抗HIV-1p24蛋白的特异性抗体。结论重组痘病毒载体rM-GP能稳定高效表达HIV-1Gag-Pol蛋白,并能引起有效的体液免疫应答。     

白藜芦醇制备方法的研究进展

综述了白藜芦醇的制备方法,其中包括植物提取法、生物技术法以及化学合成法,全面介绍了现代提取技术在白藜芦醇提取中的应用。同时对基因重组技术、细胞培养技术在白藜芦醇制备方面的应用作了系统阐述,并对其应用前景进行了评价。     

双聚类的关联规则挖掘方法

为了使所有关联规则算法都可用于双聚类挖掘,将双聚类问题转化为关联规则的频繁集挖掘问题.在为双聚类挖掘提供大量算法的同时,不但能获得双聚类,而且还能得到额外的双聚类关联信息.基因表达数据的实验结果证明了其有效性.     

Expression of green fluoscrescent protein gene in Sclerotinia sclerotiorum

Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences. The pPGF plasmid was introduced by PEG/CaCl2 treatment. Positive transformants were harvested with hygromycin B (HYG) resistance as selective marker,and then were observed with green fluorescence phenomena in response to blue light,which suggested that GFP gene was cloned into genome DNA of S. sclerotiorum. The transformants were verified mitotically stable by Southern blotting analysis and passage culturing. This study is developed as an initial step for further research into infection mechanisms of S. sclerotiorum to plants and interactions with bio-control fungus.     

Isolation and identification of a sulfate reducing bacteria and sequence analysis of its dissimilatory sulfite reductase gene

A sulfate reducing bacteria was isolated from mining sewage of Daqing Oilfield by Hungate anaerobic technology. Physiological-biochemical analysis showed that the strain could utilize polyacrylamide as sole carbon and nitrogen source. The sequence analysis of 16S rDNA illustrated that the similarity of F8 and Desulfovibrio desulfuricans (AF192153) was 99%, and the similarity sequence of dissimilatory sulfite reductase gene (DSR) cloned from the strain and Desulfovibrio desulfuricans (AF273034) was 98%. Their phylogenitic analysis was basically anastomosed, and thus temporarily named as Desulfovibrio desulfuricans F8. The DSR cloned from F8 strain was 2740 bp in length consisting of three ORF, DSRA, DSRB and DSRD as a single operon (DSRABD) regulated by the same operator. DSRA contained typical conservative box of sulfate—sulfite reducing enzyme (SiteⅠand SiteⅡ), which could bind siroheme and [Fe4S4]. DSRB retained a [Fe4S4] binding site, with an uncomplimentary structure for siroheme binding. There was no conservative box in DSRD. Sequence analysis of DSR will provide a theoretical basis for quantitative detection, metabolic pathway modification through gene engineering, and sulfate reducing bacteria (SRB) suppression.     

两种不同选择标记的转化效率比较

为了弄清两个选择标记在相同条件下的转化效率,为转基因选择遗传标记时提供参考,以优良恢复系明恢63为受体材料,采用农杆菌介导的遗传转化法,分别以hpt和bar基因为选择标记进行了转化．结果显示：1500块胚性愈伤共培养后获得214块抗性愈伤,转化效率为14．3％;5400块胚性愈伤共培养后获得156块抗性愈伤,转化效率为2．9％;两者的分化率分别为79．0％和80．7％．此外,以hpt和bar为选择标记,从胚性愈伤诱导到获得转基因植株分别需要75天和125天．因此,以hpt为选择标记的遗传转化,比较适合基因功能验证和共培养培育无选择标记水稻植株;如果是培育商品化的转基因水稻植株,则最好以bar为选择标记基因．     

微囊藻毒素生物合成研究进展

全球的淡水湖泊不同程度发生水华现象,其危害主要表现在毒素特别是微囊藻毒素的释放。有效控制微囊藻毒素应该首先了解其生物合成过程。针对这个问题,综述了微囊藻毒素的硫模板合成机制、基因簇结构、基因表达调控及环境因素对藻毒素合成的影响等方面的最新研究进展。     

免疫聚类算法在基因表达数据分析中的应用

提出了基于免疫聚类算法的基因表达数据分析方法. 根据基因表达数据矩阵的特点,设计了改进的Consine系数来度量基因相似度;借鉴生物免疫学的有关免疫理论,利用基因表达数据分析的先验知识自适应地改变抗体本身及其与抗原亲合度的关系,构造了基于免疫优势克隆的聚类算法. 与K均值算法和遗传算法的对比实验表明,该算法能够获得较大的类内紧制度、较小的类间分离度,具有较好的工程应用价值.     

基于多样化进化策略的基因表达式编程算法

针对传统GEP（Gene Expression Programming）算法的未成熟收敛以及陷入局部最优问题,提出一种基于多样化进化策略的基因表达式编程算法（DS-GEP：Gene Expression Programming based on diversified develop-ment strategy）。该算法通过基因空间均匀分布策略,自适应地交叉和变异算子以及淘汰算子等方法,对种群给予不同的进化策略,以保持种群的多样性,从而增强算法的寻优能力。通过对函数挖掘的实验证明,多样化进化策略各个部分均对改善挖掘效率发挥了作用,提高了DS-GEP函数挖掘算法的成功率。与传统GEP算法相比较,该算法的平均成功进化代数缩短了11%,成功进化时间缩短了8%,进化成功率提高了20%。