一种新的等温单向生长基因合成法初步探索

基因合成正日益成为基因获取的一种有效手段。两种常用的基因合成方法是基于PCR的基因合成（P法）和基于连接酶的基因合成(L法)。这两种方法都不是独立于所合成基因的序列内容的。文章首次提出了一种新的基因合成策略—等温单向生长法(Isothermal Unidirectional Elongation Method, IUEM), 在3种酶的作用下, DNA链在等温状态下单向串行延伸, 直到获得目标基因。由于引物设计成特殊的发夹结构, 该方法可以做到合成过程独立于或部分独立于目标基因的序列内容。本文探索了该策略的实验可行性, 检测了影响基因合成的各种因素, 并利用该方法成功地合成了一段254 bp和一段300 bp的DNA序列。     

家蚕30 kD特异性变应原的分析、纯化与质谱鉴定

以Coca's提取液分别提取到不同时期家蚕Bombyx mori的粗浸液,利用SDS-PAGE和Western blotting鉴定其特异性变应原,然后用DEAE-52离子交换层析及切胶纯化出30 kD的特异性变应原,再经MALDI-TOF在线联机分析,所得质谱数据进入网站搜索分析。结果显示：1～5龄家蚕均有20条左右蛋白带,其中 5龄家蚕有23条蛋白带,主带有11条（82、79、60、51、46、38、32、30、28、24和18 kD）。选用家蚕过敏患者阳性血清进行免疫印迹,1～4龄家蚕均显示出82和79 kD的特异性变应原;但只有5龄家蚕的30 kD蛋白为特异性变应原,通过离子交换层析和经切胶纯化出30 kD蛋白,再经MALDI-TOF-MS鉴定该蛋白为外膜蛋白。提示家蚕不同时期抗原成分有所变化,5龄家蚕新出现的30 kD蛋白为特异性变应原。     

家蚕化学诱变剂及诱导突变体的筛选

在家蚕Bombyx mori基因组计划完成之后,其功能基因组研究成为该领域的最重要课题。突变体是功能基因组学研究的重要材料,因此,通过人为诱导获取大量的突变系统是及其重要的手段。本研究用化学诱变剂ENU、MNU、DES、5-BU、EMS诱导处理家蚕标准品种C108,筛选获得了非滞育红卵,长圆筒茧、小茧、丝胶茧及绵茧突变体,致死突变体及无鳞毛蛾翅突变体。结果还表明： MNU、DES诱变家蚕的突变效率高,注射翅原基比腹部更方便且效果好,化学诱导雄体比雌体的效果好; 在时期上,注射蛹和蛾都有诱导效果。上述突变体大多为致死性突变,推测其可能与致死性基因突变有关。同时,本研究为应用TILLING技术鉴定家蚕更多目的基因突变体提供了有效的材料。     

卵形鲳鯵对刺激隐核虫的免疫应答和免疫保护研究

用刺激隐核虫(Cryptocaryon irritans)的幼虫对卵形鲳鯵(Trachinotus ovatus)进行腹腔注射和体表感染,然后每隔一周用阻动试验(Immobilization assay)检测免疫鱼的抗血清和皮肤培养液对刺激隐核虫幼虫的阻动效价,在第14周中,分别用亚致死剂量和致死剂量的刺激隐核虫幼虫对免疫鱼攻毒以检测所产生的免疫保护力。实验结果显示:两种免疫方法都能让卵形鲳鯵的血清和皮肤生成阻动刺激隐核虫幼虫的特异性抗体,并能使被免疫鱼获得明显的免疫保护,但是体表感染免疫组的血清和皮肤培养液的阻动效价都要比腹腔注射免疫组高,所获得的免疫保护力也更强。同时还发现,免疫鱼血清和皮肤培养液中的抗体存在明显的差异:两者的最初生成时间、达到峰值的时间、变化规律以及阻动效价等都不一致。因此,我们推测鱼类的系统免疫应答和皮肤黏膜免疫应答有可能是相互独立的,或者是不同步的。鱼类的体液免疫应答,特别是黏膜免疫应答对抵御刺激隐核虫的感染起了重要的作用,采用刺激隐核虫虫体疫苗可能成为预防海水鱼类白点病的一种选择。       

Leaf anatomy of Camellia sect. Oleifera and sect. Paracamellia (Theaceae) with reference to their taxonomic significance

Leaf anatomy of 21 species representing sect. Oleifera H. T. Chang and sect. Paracamellia Sealy in the genus Camellia was investigated using light microscopy. All the shapes of leaf epidermal cells were polygo-nal, and the patterns of anticlinal walls were sinuolate or sinuous, which were important to the classification of these two sections. The stomata were cyclocytic type and only present on the abaxial epidermis in both sampled sections. Seventeen leaf anatomical and cytologic characters were selected and used for the cluster analysis and principle components analyses performed by PAST program. Congruent results were obtained, with sect. Oleifera and sect. Paracamellia consistently grouping in two different clades, and some related species nesting together. This study provided leaf anatomical evidence for the merge of C. lanceoleosa H. T. Chang & J. S. Chiu and C. fluviatilis Hand.-Mazz.; of C. brevistyla Coh. St., C. obtusifolia H. T. Chang, and C. puniceiflora H. T. Chang; and of C. grijsii Hance and C. shensiensis H. T. Chang. In addition, it was also proved that C. tenii Sealy should be segregated from sect. Paracamellia and placed in the sect. Corallina Sealy. For further interest, we suggested that C. phaeoclada H. T. Chang be recognized as a subspecies of C. microphylla (Merr.) Chien. Overall, the results presented provided new insights into the relationships within a number of Camellia plants and sug-gested directions for future studies.     

Phylogenetic analysis of Paeonia sect. Moutan (Paeoniaceae) based on multiple DNA fragments and morphological data

Tree peony, being crowned the title “King of Flowers” in China, is of great medicinal, ornamental, and economic values. In the present study, the phylogeny of the wild tree peony species (section Moutan, Paeonia, Paeoniaceae), represented by twelve accessions collected from all eight species in the section, was investigated based on the DNA sequence in five DNA fragments from both nuclear (Adh1A, Adh2 and GPAT) and chloroplast (trnS-trnG and rps16-trnQ) genomes, as well as morphological characters. Both maximum parsimony (MP) and Bayesian inference of phylogeny (BI) trees were reconstructed based on the combined data of the DNA sequences and morphological data, respectively. The MP and BI trees have the similar topology, and the sect. Moutan clearly branched into two clades. One clade consists of two species, P. delavayi and P. ludlowii, corresponding to the subsect. Delavayanae, and another clade is composed of other six species. Within the second clade, the six species can be divided into three subclades consisting of P. rockii and P. decomposita, P. jishanensis and P. qiui, P. suffruticosa and P. ostii, respectively. Among the three subclades, P. jishanensis/P. qiui is most closely related to P. suffruticosa/P. ostii. These results provide up to date the clearest picture of the phylogeny of wild tree peony species in the sect. Moutan.     

极具有应用前景的生物学检测技术-生物传感器

生物传感器是近年逐渐发展起来的一种高新生物学分析检测技术,它将生物学或仿生学信号感应部件紧密连接或整合到传感系统内,具有特异、敏感、快速、便携以及操作简便等优点,发展非常迅速,并且被应用到医疗保健、食品工业、畜牧兽医等多个领域,已成为人们研究的热点之一。本文概述了生物传感器的概念与工作原理、分类、与主要领域的研究应用,分析了生物传感器的产业现状,优点与现存问题,并对其应用发展前景进行了展望。     

混合暴露条件下近江牡蛎对重金属的积累与释放特征

选择近江牡蛎作为试验生物,研究了混合暴露条件下8种重金属在近江牡蛎体内的积累和释放特征.结果表明: 近江牡蛎对重金属Pb、Cu、Ni、Cd、Cr和Hg有很强的累积能力,可较好地指示溶液中的重金属浓度水平,但对重金属Zn和As的积累能力很小,不能真实反映溶液中重金属Zn和As含量的变化水平.在随后35 d的释放阶段,8种重金属在近江牡蛎体内的含量没有明显变化,表明近江牡蛎对重金属的释放能力较差.双箱动力学模型可较好地反映混合暴露条件下近江牡蛎对重金属的积累特征,但不适合对其释放特征进行描述.     

猫尾射的生药鉴别

猫尾射是闽台民间传统的膳用中草药。本文对猫尾射的植物形态、生药性状、显微特征(包括根横切面、粉末装片)进行系统观察和理化鉴别,为其生药鉴定提供参考。     

长期施肥对黄土高原旱地黑垆土水稳性团聚体的影响

通过27年的长期定位试验,研究了长期施肥对黄土高原旱地黑垆土水稳性团聚体的影响.结果表明：与自然土壤相比,黑垆土开垦后,0～10 cm和10～20 cm土层＞0.25 mm的水稳性团聚体含量分别下降了21.35%和38.82%;0.5 mm以上各粒级的水稳性团聚体含量均呈下降趋势,其中>5 mm和5～2 mm大粒径团聚体的下降幅度最大;而0.5～0.25 mm的水稳性团聚体含量却增加了104.75%和23.13%.0～10 cm土层各粒级水稳性团聚体含量均高于10～20 cm土层.单施有机肥和有机无机肥配施处理5～2 mm粒级的水稳性团聚体含量增加最多,>5 mm粒级次之;增施有机肥、有机无机肥配合施用和秸秆还田处理能显著增加大粒径水稳性团聚体含量,改善土壤团聚体的结构.各处理0～10 cm土壤水稳性团聚体平均质量直径（MMD）均大于10～20 cm土层;自然土壤的团聚体MMD显著高于同层次耕作土壤,长期施肥处理MMD大于无肥对照,有机物质培肥处理明显高于化肥处理.土壤团聚体MMD与＞0,25 mm水稳性团聚体含量呈显著正相关.