cDNA microarray technology and its applications

The cDNA microarray is the most powerful tool for studying gene expression in many different organisms. It has been successfully applied to the simultaneous expression of many thousands of genes and to large-scale gene discovery, as well as polymorphism screening and mapping of genomic DNA clones. It is a high throughput, highly parallel RNA expression assay technique that permits quantitative analysis of RNAs transcribed from both known and unknown genes. This technique provides diagnostic fingerprints by comparing gene expression patterns in normal and pathological cells, and because it can simultaneously track expression levels of many genes, it provides a source of operational context for inference and predication about complex cell control systems. This review describes this recently developed cDNA microarray technology and its application to gene discovery and expression, and to diagnostics for certain diseases.     

Theoretical electrical conductivity of hydrogen-bonded benzamide-derived molecules and single DNA bases

A benzamide molecule is used as a “reader” molecule to form hydrogen bonds with five single DNA bases, i.e., four normal single DNA bases A,T,C,G and one for 5methylC. The whole molecule is then attached to the gold surface so that a meta-molecule junction is formed. We calculate the transmission function and conductance for the five metal–molecule systems, with the implementation of density functional theory-based non-equilibrium Green function method. Our results show that each DNA base exhibits a unique conductance and most of them are on the pS level. The distinguishable conductance of each DNA base provides a way for the fast sequencing of DNA. We also investigate the dependence of conductivity of such a metal–molecule system on the hydrogen bond length between the “reader” molecule and DNA base, which shows that conductance follows an exponential decay as the hydrogen bond length increases, i.e., the conductivity is highly sensitive to the change in hydrogen bond length.     

Eco-physiological responses of the declining population Spartina anglica to N and P fertilizer addition

Nitrogen and phosphorus are both important life elements. N, P and combined N-P fertilizers were added to the declining population Spartina anglica Hubbard in coastal China. Some growth parameters and eco-physiological responses of S. anglica to different fertilizer treatments (N, P and combined N-P fertilizer addition with high, medium and low levels, respectively) were measured. The fertilizer addition had a highly significant effect on the dynamics of its height-growth, number of leaves, number of roots and total biomass. Only N addition had a significant effect on leaf area and leaf thickness in all fertilizer treatments. On the dynamics of its height-growth, the effect of N addition was the most apparent, and the effect of N-P addition was not greater than those of N and P addition separately. The photosynthesis rate was enhanced and the yield was the highest with the highest N, the highest N-P and the medium P addition. The rates were higher than those of CK by 19.08 μmol·m^?2·s^?1, 15.47 μmol·m^?2·s^?1 and 11.23 μmol·m^?2·s^?1, respectively. The activity of SOD and POD increased with the treatments after freshwater stress for 14 days. Effects of medium N and P addition were significant for SOD activity. However, POD activity was significantly higher with the treatment of higher N and higher N-P addition. In a word, fertilizer addition improved the growth of the declining population S. anglica. The results indicated that the decline of S. anglica was correlated with the nutriment deficiency in soil, especially with the lack of N.     

Delayed development of the whitefly (Bemisia tabaci) and increased parasitism by Encarsia bimaculata in response to sublethal doses of piperonyl butoxide

Abstract Effects of sublethal piperonyl butoxide (PB) on parasitization of Bemisia tabaci (Gennadius ) (Hemiptera: Aleyrodidae) by Encarsia bimaculata Heraty et Polaszek (Hymenoptera: Aphelinidae) were evaluated both in cage and greenhouse experiments. When first, second and third instar B. tabaci nymphs were treated with PB, all but the first instar were significantly prolonged. Data indicated that sublethal PB could improve E. bimaculata parasitism rates without influencing parasitoid eclosion rates. Prolonged development increased rates of parasitism by E. bimaculata, from 17.6% to 24.7% in cages, presumably by increasing the duration of host exposure. Sublethal PB combined with E. bimaculata as an integrated approach to control B. tabaci was evaluated using life table parameters under greenhouse conditions. Indices of population trend (I) calculated from life tables were estimated at 4.6 for B. tabaci exposed to PB and parasitoids compared to 14.1 with parasitoids alone and 23.5 in untreated controls. The results showed that after PB was sprayed and parasitoids introduced, development of B. tabaci was delayed and the peak of each stage was postponed. The older nymphal stage had highest mortality, primarily due to mortality caused by parasitism by E. bimaculata.     

哈尔滨二龙山水库浮游植物及水体污染状况的研究

报道了2000 年1 月、5 月、8 月、10 月四次对哈尔滨市境内的二龙山水库的浮游植物的调查结果。共发现浮游植物46 属55 种, 其中种类最多的是绿藻和硅藻。水库四季的平均浮游藻类数量为438.46 万个/升, 绿藻占46.98%, 硅藻占44.74%, 隐藻占6.76%。优势种是平裂藻、小环藻、直链藻、针杆藻、纤维藻、空星藻、十字藻、蹄形藻、栅藻、隐藻等。对8 月份水库及上游河流的水质进行了监测和分析。根据上述结果对水质进行了评价, 并对二龙山库区的水土保持及可持续性发展进行了探讨。     

白花蛇舌草醇提取物对乳腺癌T-47D细胞株生长的抑制作用

目的：观察白花蛇舌草醇提取物对乳腺癌细胞T-47D生长的影响。方法：采用平皿克隆形成实验、软琼脂集落形成实验和BrdUrd（溴脱氧尿苷）掺入实验,观察不同浓度的白花蛇舌草醇提取物（1.0μg/mL,3.0μg/mL,10μg/mL,30μg/mL,100μg/mL）对T-47D细胞锚定依赖性生长和软琼脂集落形成能力及核酸合成的抑制作用。结果：①随着白花蛇舌草醇提取物浓度的升高,对T-47D细胞的生长呈现明显抑制作用;②白花蛇舌草醇提取物可呈浓度依赖性抑制对数生长期T-47D细胞核酸合成。结论：白花蛇舌草醇提取物可能通过影响肿瘤细胞核酸合成而抑制T-47D细胞生长。     

“两广二号”家蚕抗病毒基因Bmlipase-1和BmSP-2克隆与原核表达

本研究以优良杂交品种"两广二号"家蚕为试材,克隆了该杂交品种家蚕两个抗家蚕核型多角体病毒(BmNPV)基因:脂肪酶基因Bmlipase-1和丝氨酸蛋白酶基因BmSP-2,测序并分别与不同品种蚕的同源基因序列进行比较。结果显示,"两广二号"家蚕Bmlipase-1基因ORF长度为885bp,编码294个氨基酸,BmSP-2扩增长度为855bp,编码284个氨基酸;它们的核苷酸和推导氨基酸序列同源性皆达92%以上,Bmlipase-1更保守,同源性大于99%";两广二号"家蚕的Bmlipase-1基因脂肪酶活化部位和BmSP-2基因酶催化三联体位点的氨基酸残基与不同品种蚕的完全相同。以上结果说明这两个抗病毒基因在蚕的遗传进化过程中高度保守,提示其可能在机体消化或者免疫防御方面起着重要生理作用。将这两个抗病毒基因在大肠杆菌BL21中进行融合表达,获得的融合Bmlipase-1和BmSP-2蛋白分子量分别为47kD和42kD左右。     

蚜科专化菌努利虫疠霉菌种超低温储存

虫霉目真菌的活力对超低温储存较为敏感。储存法在大范围应用前,需对储存效果进行详细评估。将蚜科专化菌努利虫疠霉以初级分生孢子形式（2－3′105个孢子/mL）在-80℃超低温存储12个月。结果显示日常用于培养该真菌的含0.1%乳化芝麻油的萨氏培养基作为超低温保护基质能有效地储存努利虫疠霉孢子,比常见的冷冻保护剂如二甲亚砜和甘油的效果好。孢子悬液经解冻和培养后可获得最多的生物量,而且菌种保持了较高的生长速率。更重要的是,萨氏培养基的主要成分4%葡萄糖、1%蛋白胨和1%酵母粉在低温存储过程中发挥了协同作用,能保     

F43Y及I354M,L358F定点突变对植酸酶热稳定性及酶活性的改善

对重组酵母PPNPm8的植酸酶phyAm基因进行PCR介导的定点突变,即将植酸酶43位的苯丙氨酸替换为酪氨酸(F43Y),将其354、358位的异亮氨酸、亮氨酸分别替换为甲硫氨酸和苯丙氨酸(I354M,L358F),得到了2个突变体PPNPm-1(F43Y)及PPNPm-2(I354M,L358F).含突变基因的重组表达载体pPIC9kphyAm-1,pPIC9kphyAm-2在毕赤酵母GS115中表达,对表达产物进行酶活性测定及热稳定性检测.结果表明:突变体PPNPm-1最适反应温度比未突变体PPNPm8上升了3℃,75℃处理10min,热稳定性提高15%,比活力提高11%;PPNPm-2最适反应温度未改变,热稳定性比PPNPm8仅提高3%,比活力降低6.5%.对突变前后的植酸酶空间结构进行比较预测,发现突变氨基酸Tyr43与空间位置相邻的Asn416之间形成氢键,增强了酶的热稳定性.