微量元素对动物基因表达的调控

随着营养学和现代生物技术的发展,发现日粮中微量元素在一定程度上影响动物基因的表达,这种作用可以发生在转录水平,也可以发生在转录后水平,进而影响机体的整个代谢过程.综述微量元素对基因表达调控的主要机制、存在方式,并介绍铁、硒、锌、铜、铅等元素对部分基因表达的主要影响.     

湖南石门雄黄矿区矿样中的砷氧化细菌的分离及鉴定

绿色的生物学方法处理含砷废水,对于缓解工业发展所带来砷污染问题,提高居民健康生活质量具有重要意义。以湖南石门雄黄矿区的尾矿池积水（SY）、地下400 m处的矿坑水（NY）和尾矿池底泥表层（XY）3个样本为研究对象,利用细菌富集和多次传代的方法,从这3个环境样本中获得可氧化三价砷的3个菌群。在砷质量浓度为l~5 g·L-1的培养基中分别检测上述3个菌群的耐受能力,发现在砷质量浓度高达5 g·L-1的条件下,上述菌群均能在2~3 d内达到109 cells·mL-1。利用上述菌群处理l g·L-1亚砷酸钠溶液,SY、XY和NY菌群可分别在25、20和35 h内将三价砷完全氧化,并且使溶液中的总砷质量浓度降低66.7%。进一步对3个菌群进行多次平板分离,利用五价砷和硝酸银的颜色反应,获得11株可氧化三价砷的目的菌株。对这些菌株16S rDNA进行NCBI的BLASTN序列比对发现,菌株SMY24、SMY33、SMY22、SMY32、SMY21和SMY31均属于假单胞菌属（Pseudomonas）,菌株SMY104属于不动杆菌属（Acinetobacter）,菌株SMY17、SMY25、SMY9和SMY1在所有的已知序列中,最大的相似率只有91%,最小的相似率为76%,且都是未曾报道纯培养的细菌。对这些菌株三价砷氧化能力进行测定发现,其中菌株SMY104和SMY21在20 h内能将三价砷几乎完全氧化。通过对这些菌株三价砷氧化酶基因进行初步检测,发现了多个菌株具有三价砷氧化酶基因（AoxB）。     

Cloning and expression analysis of geosmin synthase gene from Aphanizomenon gracile北大核心CSCD

Geosmin is a secondary metabolite responsible for earthy odors. The occurrence of geosmin has great impact on the quality of water environment. The gene essential for geosmin biosynthesis have been identified in several species. But little is known about the mechanism of geosmin synthesis in Aphanizomenon gracile. This study attempted to clone the gene involved in geosmin biosynthesis of Aphanizomenon gracile a nd a nalyze t he geosmin production u nder d ifferent e nvironments. T he high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) was used to amplify the full-length of geosmin synthase gene from Aphanizomenon gracile (WH-1). Real time PCR (RT-PCR) was applied to quantify the geosmin production in different light and temperature. As a result, geo, a geosmin synthase gene from Aphanizomenon gracile (WH-1) was cloned by hiTAIL-PCR. The full-length open reading frame (ORF) of geo was 2 262 bp, coding for a protein of 753 amino acids. Meanwhile, WH-1 was treated with different environment conditions and mRNA expression levels of geo were quantified by RT-PCR. It was found that low temperature (15 °C), high light intensity (35 μmol m-2 s-1) and continuous light illumination were beneficial to the expression of geo. The successful amplification of geosmin synthase gene verified that hiTAIL-PCR is an effective and simple procedure of low cost. The result provides fundamental knowledge on the monitoring and prevention for odorants.     

Interspecific and intraspecific identification of Dendrobium based on the psbAtrnH intergenic region sequences and the 5S rRNA gene spacer sequences北大核心CSCD

This research aimed to investigate the interspecific and intraspecific identification of Dendrobium by using the multi-locus method so as to provide a molecular basis for Dendrobium identification through the combination of chloroplast psbA-trnH intergenic region sequences and ribosome 5S rRNA gene spacer sequences. PCR direct sequencing was applied to detect the chloroplast psbA-trnH intergenic region sequences as well as the ribosome 5S rRNA gene spacer sequences of 12 Dendrobium species, while the psbA-trnH intergenic region sequences of Dendrobium denneanum dq-2 variety and dq- 5line were cloned and sequenced for single nucleotide polymorphism (SNP) analyzing. The sequences were analyzed by the software Sequencher4.14, Bioedit7.0, MEGA5.2 and Dansp5.0; the interspecific and intraspecific Kimara-2-Parameter(K2P) distances were also calculated. The phylogenetic tree (using Neighbor joining method) was constructed with Bulbophyllum odoratissimum and Bletilla striata as outgroup. The results showed an average length of chloroplast psbA-trnH gene sequences in Dendrobium as 742.3 bp, with 72 variable sites, including 33 information sites; the average length of the ribosome 5S rRNA gene spacer sequences in Dendrobium was 336.4 bp, with 213 variable sites including 139 information sites. Using psbAtrnH intergenic region sequences in combination with ribosome 5S rRNA gene spacer sequences can not only identify D. denneanum, D. hancockil, D. thysiflorum, D. devonianum, D. moniliforme, D. chrysotoxum, D. officinale, D. heterocarpum and D. nobile, but also differentiate D. officinale from different geographical populations, and distinguish the dq-2 variety and dq 5line with SNP in the multi locus of D. denneanum.     

基因枪介导获得转植酸酶基因水稻研究

植酸酶是一类能促进植酸及其盐类水解成肌醇和磷酸的酶的总称.它能提高饲料和食品中磷的利用率,减轻粪便中磷排放所造成的环境污染,清除植酸与金属离子的螯合作用,改善微量元素的吸收和利用.本研究将马铃薯的胞外分泌信号肽(Psec)和大肠杆菌的植酸酶基因(appA2)结合组成融合基因导入粳稻台北309中.PCR检测和Southern杂交表明,植酸酶基因和分泌信号肽序列已经整合到水稻的基因组中.无机磷含量分析表明,含目的基因的转基因水稻种子及茎叶中的无机磷含量均较未转化植株有显著的提高.图6表1参14     

肺炎嗜衣原体主要外膜蛋白基因片段在大肠杆菌中的表达与鉴定

表达肺炎嗜衣原体（Chlamydia pneumoniae,Cpn）主要外膜蛋白（MOMP）的可变区VD2-VD3区．纯化产物并进行免疫活性分析,为探索重组蛋白在肺炎嗜衣原体血清学诊断中的应用提供资料．应用聚合酶联反应（PCR）技术,从肺炎嗜衣原体标准株AR-39的MOMP上扩增出抗原优势表住VD2-VD3区,将目的片段定向插入pET-30a载体,转化大肠杆菌B121,IPTG诱导表达并以Ni-NTA亲和层析柱纯化表达产物并行western-blot鉴定．成功构建了pET-30a-MOMPVD2-VD3的原核表达系统,表达并纯化出相对分子质量（Mr）为24×10^3Da的重组蛋白．Western-blot证实重组蛋白能与Cpn MOMP多克隆抗体发生特异性反应．肺炎嗜衣原体的MOMPVD2-VD3基因可以在大肠杆菌中得到表达,其表达产物能与相应的抗体结合,为肺炎嗜衣原体诊断候选抗原的研究奠定了基础．图4,参9．     

Validation of Bacteroidales-based microbial source tracking markers for pig fecal pollution and their application in two rivers of North China

• Pig feces is the predominant excrement produced by animal husbandry in China. • The PF, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan MST assays showed better performance. • The pig-specific MST assays can contribute to managing the pig fecal pollution. In China, pig feces is the predominant source of excrement produced by animal husbandry. Improper use or direct discharge of pig feces can result in contamination of natural water systems. Microbial source tracking (MST) technology can identify the sources of fecal pollution in environmental water, and contribute to the management of pig fecal pollution by local environmental protection agencies. However, the accuracy of such assays can be context-dependent, and they have not been comprehensively evaluated under Chinese conditions. We aimed to compare the performance of five previously reported pig-specific MST assays (PF, Pig-Bac1^SYBR, Pig-Bac2^SYBR, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan, which are based on Bacteroidales 16S rRNA gene markers) and apply them in two rivers of North China. We collected a total of 173 fecal samples from pigs, cows, goats, chickens, humans, and horses across China. The PF assay optimized in this study showed outstanding qualitative performance and achieved 100% specificity and sensitivity. However, the two SYBR green qPCR assays (Pig-Bac1^SYBR and Pig-Bac2^SYBR) cross-reacted with most non-pig fecal samples. In contrast, both the Pig-1-Bac^TaqMan and Pig-2-Bac^TaqMan assays gave 100% specificity and sensitivity. Of these, the Pig-2-Bac^TaqMan assay showed higher reproducibility. Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand, Japan, and America. Using the PF and Pig-2-Bac^TaqMan assays, a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.     

Biofiltration and disinfection codetermine the bacterial antibiotic resistome in drinking water: A review and meta-analysis

• Published data was used to analyze the fate of ARGs in water treatment. • Biomass removal leads to the reduction in absolute abundance of ARGs. • Mechanism that filter biofilm maintain ARB/ARGs was summarized. • Potential BAR risks caused by biofiltration and chlorination were proposed. The bacterial antibiotic resistome (BAR) is one of the most serious contemporary medical challenges. The BAR problem in drinking water is receiving growing attention. In this study, we focused on the distribution, changes, and health risks of the BAR throughout the drinking water treatment system. We extracted the antibiotic resistance gene (ARG) data from recent publications and analyzed ARG profiles based on diversity, absolute abundance, and relative abundance. The absolute abundance of ARG was found to decrease with water treatment processes and was positively correlated with the abundance of 16S rRNA (r² = 0.963, p<0.001), indicating that the reduction of ARG concentration was accompanied by decreasing biomass. Among treatment processes, biofiltration and chlorination were discovered to play important roles in shaping the bacterial antibiotic resistome. Chlorination exhibited positive effects in controlling the diversity of ARG, while biofiltration, especially granular activated carbon filtration, increased the diversity of ARG. Both biofiltration and chlorination altered the structure of the resistome by affecting relative ARG abundance. In addition, we analyzed the mechanism behind the impact of biofiltration and chlorination on the bacterial antibiotic resistome. By intercepting influent ARG-carrying bacteria, biofilters can enrich various ARGs and maintain ARGs in biofilm. Chlorination further selects bacteria co-resistant to chlorine and antibiotics. Finally, we proposed the BAR health risks caused by biofiltration and chlorination in water treatment. To reduce potential BAR risk in drinking water, membrane filtration technology and water boiling are recommended at the point of use.     

浮游动物DNA宏条形码标志基因比较研究

DNA宏条形码技术作为一种新型生物监测方法,在未来生态环境监测中有巨大的应用潜力。目前,浮游动物DNA宏条形码监测仍在发展阶段,需要首先对其(采样方法、引物选择和数据分析等)进行标准化和调整,然后才能用于常规流域生态监测。其中,如何选择合适的PCR扩增引物是DNA宏条形码生物监测标准化的关键问题之一。本研究比较了COI、18SV9和16S通用引物在浮游动物DNA宏条形码监测中的差异,为初步建立规范化的浮游动物DNA宏条形码监测方法提供技术支撑。结果表明,16S引物对浮游动物具有更好的特异性,其产生的操作分类单元(operational taxonomic unit, OTU)有88.1%属于浮游动物。虽然18SV9引物具有更高的物种覆盖度,不仅能扩增出浮游动物,还能扩增出大量藻类和真菌,但其物种识别敏感性较差,不适合浮游动物物种水平多样性监测。COI引物的浮游动物物种特异性、物种覆盖度和物种识别敏感性都适中,检出的浮游动物物种数量高于18SV9引物和16S引物,更加适合浮游动物DNA宏条形码监测。