三倍体毛白杨及杉木苯酚液化物的结构分析

为搞清酸性条件下三倍体毛白杨和杉木苯酚液化物的结构,该研究对两种木材分别进行了液化处理,并对液化物进行了傅立叶转换红外光谱（FTIR）、核磁共振光谱（1H-NMR、13C-NMR和31P-NMR）以及电子扫描电镜（SEM）的测定分析.FTIR和NMR的分析结果表明,液化后的毛白杨和杉木均发生了结构上的明显变化,出现了纤维素和木素的基本活性结构单元,表明液化处理使木材发生了降解、酚化等化学反应.SEM测定结果表明,木材液化物中含有未完全液化的微小木材组织碎片,两种木材液化物中残存的这种物质的大小存在差异.      

苏云金芽孢杆菌杀虫晶体蛋白Cry1Ba3活性区的研究

 通过对Cry1Ba3蛋白序列的分析，以及与已知Cry蛋白比较，设计了6对特异性引物，PCR扩增获得6种不同长度的cry1Ba3基因片段。将这些基因片段克隆到pET-21b载体上，导入大肠杆菌BL21中进行诱导表达，最终得到6种不同长度的Cry1Ba3蛋白片段。采用浸叶法检测这些蛋白片段对小菜蛾的杀虫活性，研究结果表明含有第1-685位和含有第22-655位氨基酸的蛋白片段对小菜蛾的毒性，与全长Cry1Ba3蛋白相比，没有改变；含有第54-655位氨基酸的蛋白片段对小菜蛾的毒力明显降低；而含有第22-627位和含有第85-655位氨基酸的蛋白片段完全丧失了对小菜蛾的活性。上述结果表明Cry1Ba3蛋白对小菜蛾的最小活性区在第22-655位氨基酸之间。     

The Minimal Active Fragment of the Cry1Ai Toxin is Located Between 36I and 605I

The novel cry1Ai gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of Cry1Ai. All truncated fragments were expressed in Escherichia coli strain BL21 (DE3), and the insecticidal activity against 2nd- instar P. xylostella larvae was assessed using full-length Cry1Ai as a positive control. The results indicate that the minimal active fragment of the Cry1Ai toxin against P. xylostella is located between amino acid residues 36I and 605I, which is smaller than the regions previously reported for Cry1A. The first two amino acids (34T and 35P) on helix α-1 and whole helix α-2 of domain I and sheet β-32 of domain III are necessary for Cry1Ai toxin to keep its toxicity against P. xylostella.     

苦楝果活性成分及其地理变异研究进展

苦楝(Melia azedarach Linn.)在我国分布广泛,地理变异明显,生长迅速,材质优良,是良好的用材树种;其根、皮、花、果皆可入药,对多种害虫有明显的毒杀、拒食和忌避活性,是常见的生物农药原料,将苦楝果作为活性成分提取材料,有利于开发材用价值.文章综述了苦楝果提取物的抗虫作用、杀虫活性成分及其作用效果,并结合遗传育种策略,针对国内对苦楝果活性成分地理变异研究的局限与不足,提出在全分布区内收集具代表性的苦楝种源或家系,分析不同种源、家系间苦楝果的活性成分种类及含量差异,研究其地理变异及个体变异规律,在优良种源或家系选择的基础上,进一步将苦楝的生物农药应用结合木材利用进行改良,选择果实中活性成分含量高及生长性状表现优良的种源或家系,再通过遗传育种手段培育出优良品种用于果林营造,以期达到苦楝材用与药用相结果的目的,综合开发苦楝的经济价值.     

Pemoline and magnesium hydroxide: lack of effect on RNA and protein synthesis

Brain RNA polymerase isolated from rats treated with pemoline and magnesium hydroxide (Cylert) was not more active than enzyme from control animals. The drug did not increase enzymic activity in vitro. Pemoline did not significantly affect either RNA or protein synthesis in suspensions of Ehrlich ascites carcinoma cells.     

香蕉枯萎病内生生防菌H4-3抑菌蛋白稳定性研究

铜绿假单胞菌H4-3菌株是福建省农科院农业生物资源研究所微生物研究中心从香蕉植株中分离到的内生菌,该菌株对香蕉枯萎病原菌具有很强的抑菌活性,其活性物质为大分子蛋白.研究测定了H4-3菌株抑菌蛋白的稳定性,结果表明：H4-3菌株的抑菌蛋白耐高温、强酸、强碱和高浓度蛋白酶,紫外线和反复冻融对H4-3菌株抑菌蛋白活性无显著影响.     

家蚕血液中超氧物岐化酶（SOD）的研究

用连苯三酚自氧化法测定家蚕血液中超氧物岐化酶活性的结果表明，家蚕血液中超氧物岐化酶活性不仅在品种间存在差异，而且在同一品种的雌雄体间亦存在明显差异，不同发育期其血液中超氧物岐化酶的活性和同功酶均有差异．幼虫期同一品种雄蚕血液中超氧物岐化酶活性高于同一生长期的雌蚕，五龄蚕超氧物岐化酶活性低于四龄蚕．幼虫期与蛹期超氧物岐化酶聚丙烯酰胺凝胶电泳显示出活性谱带亦不相同，幼虫期多为一条活性带，且活性带峰值是雄蚕高于雌蚕；而蛹期有的品种出现两条活性带，其峰值则显示出雌蚕高于雄蚕     

用cDNA-AFLP技术研究茶树种子在低温贮藏过程中差异基因的表达

以茶树无性系龙井43种子为材料,利用cDNA-AFLP差异显示方法,分析它在低温贮藏后的基因表达,为顽拗性植物种子的中长期保存提供理论依据。从64对引物筛选出的差异片段中克隆出10个低温差异表达片段,有7个在GenBank数据库中与小分子量热激蛋白、MYB类转录因子、衰老相关蛋白、F-box家族蛋白、蛋白激酶、磷酸二酯酶等基因有较高同源性。     

Bovine growth hormone: human food safety evaluation

Scientists in the Food and Drug Administration (FDA), after reviewing the scientific literature and evaluating studies conducted by pharmaceutical companies, have concluded that the use of recombinant bovine growth hormone (rbGH) in dairy cattle presents no increased health risk to consumers. Bovine GH is not biologically active in humans, and oral toxicity studies have demonstrated that rbGH is not orally active in rats, a species responsive to parenterally administered bGH. Recombinant bGH treatment produces an increase in the concentration of insulin-like growth factor-I (IGF-I) in cow's milk. However, oral toxicity studies have shown that bovine IGF-I lacks oral activity in rats. Additionally, the concentration of IGF-I in milk of rbGH-treated cows is within the normal physiological range found in human breast milk, and IGF-I is denatured under conditions used to process cow's milk for infant formula. On the basis of estimates of the amount of protein absorbed intact in humans and the concentration of IGF-I in cow's milk during rbGH treatment, biologically significant levels of intact IGF-I would not be absorbed.     

松墨天牛和中国圆田螺体内纤维素酶系的比较研究*

 分别在不同pH,温度、反应时间条件下,对松墨天牛和中国圆田螺两种动物体内纤维素酶系的组成和活性进行了分析;在纤维素酶系的3种底物同时存在下,分析了两物种中纤维素酶的协同作用。结果表明在松墨天牛和中国圆田螺体内均具有将纤维素降解为简单糖的完整纤维素酶系。松墨天牛体内纤维素酶系中各组分的活性均较中国圆田螺高;在对3种底物同时作用时,松墨天牛体内纤维素酶系活性也高于中国圆田螺,揭示了两物种的纤维素酶系的协同作用有明显差异,为进一步寻求高活性纤维素酶提供基础数据。     

查尔酮合酶蛋白表达技术、结构、活性和进化

查尔酮合酶(CHS)是类黄酮途径的首步关键酶,参与合成所有类黄酮和异黄酮物质,参与决定多种植物重要性状。NCBI已有2917条CHS序列登录和释放。单个物种基因组中的CHS普遍由基因家族编码,通过基因加倍而产生。CHS蛋白的表达技术目前均是基于大肠杆菌的原核表达,多采用Novagen公司的pET载体系统,最通行参数为37℃条件下1 mmol/L IPTG诱导3h,表达蛋白普遍采用His-Tag进行亲和纯化,采用质谱或免疫分析技术进行身份检测,通过对生化反应底物和产物的HPLC检测可以精确分析酶活。苜蓿等植物的CHS蛋白已完成X射线晶体衍射研究,获得了三维结构和活性位点构象的初步解析,并与植物Ⅲ型PKS超家族中的其它酶类进行了比较。CHS蛋白的催化活性中心含有一个Cys-His-Asn三联体,另外CoA结合通道和内部的起始/延伸/环化腔关系到CHS蛋白的底物特异性及聚酮化合物链延伸的长度。植物Ⅲ型PKS超家族中,其它酶的结构骨架和催化方式与CHS相似,但活性位点残基的变化可能会造成活性中心结构的改变,进而产生底物特异性、催化能力和产物种类的显著变化,是蛋白水平进化的根本动力。     

高体革鯻线粒体DNA16S rRNA和COI基因片段的克隆和序列分析（摘要）

[目的]为高体革鯻的遗传资源、物种间的亲缘关系和系统进化等研究提供一定的生物学依据。[方法]采用PCR扩增和序列测定等技术,对高体革鯻线粒体DNA16S rRNA和COI基因片段进行初步研究。[结果]经PCR扩增和测序,分别得到16SrRNA和COI基因片段的碱基序列,其中16SrRNA基因片段的大小为791bp,碱基A、T、G、C的含量分别为31.6%、21.4%、20.4%和26.7%;COI基因片段的大小为631bp,碱基A、T、G、C含量分别为27.7%、23.6%、29.8%和18.9%。在这2个基因片段中,GC含量均低于AT含量,AT/GC分别为1.13和1.05。[结论]通过对高体革鯻16SrRNA和COI2个基因片段遗传特征的研究,发现其种内变异比较低,在3个样本中16SrRNA基因片段序列完全一样,COI基因片段也完全一样,说明高体革鯻的这2个基因都非常保守。     

高体革鯻线粒体DNA 16S rRNA和COI基因片段的克隆与序列分析

[目的]为高体革鯻的遗传资源、物种间的亲缘关系和系统进化等研究提供一定的生物学依据。[方法]采用PCR扩增和序列测定等技术,对高体革鯻线粒体DNA 16S rRNA和COI基因片段进行初步研究。[结果]经PCR扩增和测序,分别得到16S rRNA和COI基因片段的碱基序列,其中16SrRNA基因片段的大小为791 bp,碱基A、T、G、C的含量分别为31.6%、21.4%、20.4%和26.7%;COI基因片段的大小为631 bp,碱基A、T、G、C含量分别为27.7%、23.6%、29.8%和18.9%。在这2个基因片段中,GC含量均低于AT含量,AT/GC分别为1.13和1.05。[结论]通过对高体革鯻16S rRNA和COI2个基因片段遗传特征的研究,发现其种内变异比较低,在3个样本中16S rRNA基因片段序列完全一样,COI基因片段也完全一样,说明高体革鯻的这2个基因都非常保守。     

口蹄疫病毒VP1基因在胡萝卜中的表达

为了生产一种可以直接口服的口蹄疫疫苗,以胡萝卜(Daucus carota L.var.sativa)无菌幼苗下胚轴诱导的愈伤组织为受体材料,通过农杆菌LBA4404介导的遗传转化,在CaMV双35S启动子的驱动下,将口蹄疫病毒(foot-and-mouth disease virus,FMDV)结构蛋白VP1基因的活性肽片段(包括2个141~160位肽和1个200~213位肽的基因片段)导入胡萝卜愈伤组织细胞。经PCR和PCR-Southern杂交等方法鉴定转化苗,确认口蹄疫病毒结构蛋白VP1基因已整合到胡萝卜染色体中,转化效率达到52%。SDS-PAGE试验结果初步证明,在转化苗总蛋白中有目的基因表达,胡萝卜种子中可溶性蛋白含量最高,达3.57 g/L,而根中可溶性蛋白含量最低,仅有0.17 g/L;靠近心叶的叶片可溶性蛋白含量可达到1.475 g/L,而最外部叶片可溶性蛋白含量仅有0.37 g/L,相差近5倍。Dot-ELISA和ELISA检测结果表明,转化苗中表达的口蹄疫病毒结构蛋白VP1可以与FMDV灭活疫苗免疫动物制备的抗体特异结合。以上试验结果证明,口蹄疫病毒结构蛋白VP1基因在转基因胡萝卜中表达成功。     

Phylogenetic Relationships in Genus Arachis Based on SSR and AFLP Markers

Fourteen wild species of different sections in the genus Arachis and 24 accessions of the AABB allotetraploid A. hypogaea （cultivated peanut） from several countries which belong to different botanical varieties, were analyzed by SSR and AFLP marker systems. The assay-units per system needed to distinguish among all the tested accessions were at least five for SSR or two for AFLP. The genetic distance detected by the SSR markers ranged from 0.09 to 0.95, and the mean was 0.73; and the genetic distance detected by the AFLP markers ranged from 0.01 to 0.79 with an average of 0.42. All the tested peanut SSR primer pairs were multilocus ones, and the amplified fragments per SSR marker in each peanut genome ranged from 2 to 15 with the mean of 4.77. The peanut cultivars were closely related to each other, and shared a large numbers of SSR and AFLP fragments. In contrast, the species in the genus Arachis shared few fragments. The results indicated that the cultivated peanut （A. hypogaea L.） varieties could be partitioned into two main groups and four subgroups at the molecular level, and that A. duranensis is one of the wild ancestors of A. hypogaea. The lowest genetic variation was detected between A. cardenasii and A. batizocoi, and the highest was detected between A. pintoi and the species in the section Arachis. The relationships among the botanical varieties in the cultivated peanut （A. hypogaea L.） and among wild species accessions in section Arachis and those in other sections in the genus Arachis were discussed.     

我国不同省份高蛋白栽培大豆的表型性状变异分析

利用<中国大豆品种资源目录>数据库提供的数据,对我国高蛋白材料在不同省份之间的变异规律进行了统计分析,提出了黄淮夏大豆生态区是我国的高蛋白大豆的遗传多样性中心.     

Study on Somaclonal Variation of Spring Wheat

Somaclonal variation of calli and regenerated plants of spring wheat were detected by using technique RAPD in the study. Calli at different culture stages and regenerated plants derived from young spikes and immature embryos were used as materials. Molecular variation could be reflected from electrophoresis pattern of RAPD fragments at different culture stage in calli, and in regenerated plants derived from different explants, even no phenotype variations were found. Somaclonal variation in calli and in regenerated plants appeared regularly: A higher frequency of variation in hybrids F2 was detected than that of the cultivar that is stable genetically. High variation frequency of RAPD fragments appeared in calli when cultured 75 days. The identical variations of RAPD fragments were observed in calli and in the regenerated plants induced from different genotype or explants. The variation frequency detected is higher in regenerated plants than that of in calli. RAPD could be applied easily and simply to determine variation in level of DNA at each stage cultured in vitro.     

Small DNA deletions creating avirulence in Streptococcus pyogenes

The M protein is the antigen on the surface of group A streptococci that allows these bacteria to resist phagocytosis. DNA encoding the M12 protein was cloned into Escherichia coli and used as an isotopically labeled hybridization probe to compare genomic DNA's isolated from M+ and M- isogenic cultures in an effort to elucidate the genetic basis of this variation. DNA's from two spontaneous, independent M- variants contained small (approximately 50 base pairs) deletions which were mapped to identical restriction fragments within or adjacent to the M protein coding sequence. Taken together with the pleiotropic nature of these deletions, this suggests that they define a regulatory switch.     

纳米铜对大鼠肝脏毒性相关蛋白过氧化氢酶的分离鉴定及生物信息学分析

【目的】分离和鉴定纳米铜对大鼠肝脏毒性相关蛋白过氧化氢酶(catalase, CAT),探讨CAT在毒性发挥中的作用,为揭示纳米铜对肝脏毒性机制提供依据。【方法】应用2-DE技术和PDQuest 8.0软件在大鼠肝脏蛋白组中筛选纳米铜对肝脏毒性差异蛋白,经质谱鉴定后进行生物信息学分析。【结果】筛选到下调的差异蛋白点6602和7702与肝毒性相关,鉴定均为CAT蛋白;其性质稳定,有一定亲水性,无信号肽,定位于细胞质,可能属于非分泌性蛋白,含有过氧化氢酶活性位点64FDRERIPERVVHAKGAG80和过氧化氢酶亚铁血红素配合基位点354RLFAYPDTH362等功能位点;无规则卷曲、α螺旋和延伸链是其主要的二级结构元件,并预测了其三级结构图;同源性分析表明,大鼠的CAT与其它8个物种有较高同源性,并构建了CAT 蛋白的系统进化树。【结论】纳米铜通过下调大鼠肝脏中CAT蛋白表达,引起肝细胞氧化应激损伤,可能是其发挥毒性作用的途径之一。     

Allergic encephalomyelitis: passive transfer prevented by encephalitogen

Allergic encephalomyelitis was produced in rats by passive transfer of lymph node cells from donors immunized intradernmally with nleural tissute or an encephalitogenic basic protein pluts adjulvants. The same basic protein, injected intravenously into the recipients before or after transfer of lymph node cells, prevented the disease. Even established lesions were reversed. Inhibition by basic protein was specific for encephalomyelitis; it had no effect onz passive transfer of allergic adrenalitis.