Selectable marker replacement in Saccharomyces cerevisiae

Selectable markers integrated by the ‘gamma’ deletion method (Sikorski and Hieter, 1989) can be efficiently replaced in vivo with other markers by transformation with homologous plasmids. Transformation frequencies in experiments designed to replace original selectable markers with an alternate marker were high and molecular analysis confirmed that all transformants that exhibited the expected phenotypes (loss of the original prototrophy and gain of the alternate prototrophy) resulted from homologous recombination between plasmid sequences at the target locus. This technique involves no plasmid construction and greatly facilitates the generation of yeast cells containing multiple gene disruptions.     

Early history,discovery, and expression of Aequorea green fluorescent protein,with a note on an unfinished experiment

The bioluminescent hydromedusan jellyfish, Aequorea victoria, emits a greenish light (λ_max = 508 nm) when stimulated electrically or mechanically. The light comes from photocytes located along the margin of its umbrella. The greenish light depends on two intracellular proteins working in consort: aequorin (21.4 kDa) and a green fluorescent protein (27 kDa). An excited state green fluorescent protein molecule results, which, on returning to the ground state, emits a greenish light. Similarly, a green light emission may be induced in the green fluorescent protein by exposing it to ultraviolet or blue light. Because the green light can be readily detected under a fluorescence microscope, the green fluorescent protein, tagged to a protein of interest, has been used widely as a marker to locate proteins in cells and to monitoring gene expression. This article reviews the work that took place leading to the discovery, cloning, and expression of the green fluorescent protein, with a note on an unfinished experiment. Microsc. Res. Tech. 73:785–796, 2010. © 2010 Wiley‐Liss, Inc.     

Concentrations of host-specific and generic fecal markers measured by quantitative PCR in raw sewage and fresh animal feces

We measured the concentrations of four host-specific (human, dog, cow, and horse Bacteroidales), four generic fecal (16S total Bacteroidales and Escherichia coli, 23S Enterococcus and uidA E. coli,) and two universal bacterial (16S universal and rpoB universal) DNA targets by qPCR in raw sewage and pooled fecal samples from dogs, cows, horses, and Canada Geese. A spiking protocol using the non-fecal bacterium Pseudomonas syringae pph6 was developed to estimate the recovery of DNA from fecal and environmental samples. The measured fecal marker concentrations were used to calculate baseline ratios and variability of host-specific to generic indicators for each host type. The host-specific markers were found in high concentrations (8-9 log₁₀ copies/g dry wt.) in their respective hosts' samples, which were equal to or greater than the concentrations of generic E. coli and Enterococcus markers, lending support to the use of host-specific and generic Bacteroidales as sensitive indicators of fecal pollution. The host-specific markers formed a consistent percentage of total Bacteroidales in target host feces and raw sewage, with human-specific comprising 82%, dog-specific 6%, cow-specific 4% and horse-specific 2%. Based on this limited data set, the measurement of host-specific indicators by qPCR has several promising applications. These applications include determining the percentage of total Bacteroidales contributed by a specific host type, using the ratios of host-specific markers to E. coli or Enterococcus to estimate the contribution of each source to these regulated fecal indicator bacteria, and estimating the mass of feces from each host type in environmental samples.     

5种绿藻对几种常用抗生素的敏感性

考察莱茵衣藻、亚心形扁藻、四爿藻、微绿球藻和小球藻5种绿藻对几种常用抗生素的敏感性,结果表明,莱茵衣藻对卡那霉素、四环素敏感,敏感浓度(细胞致死浓度,下同)均为25μg/mL,对氯霉素敏感性次之,而对氨苄青霉素不敏感;亚心形扁藻和四爿藻对氯霉素敏感,敏感浓度均为25μg/mL,对四环素、卡那霉素和氨苄青霉素不表现敏感性;小球藻和微绿球藻对实验中的4种抗生素都不敏感。     

Detection and analysis of quantitative trait loci affecting production and secondary traits on chromosome 7 in Israeli Holsteins

A total of 5,459 Israeli Holstein cows, daughters of 11 sires, were genotyped for 29 microsatellites spanning chromosome 7 and analyzed by the daughter design for 9 economic traits: milk, fat, and protein yield, fat and protein percentage, somatic cell score, female fertility, herd life, and milk persistency. Quantitative trait loci at chromosome-wise 0.05 significance were obtained for fat and protein yield, fat percentage, somatic cell score, and female fertility. Peak F-values were obtained at 29 cM for fat and protein yield and fat percentage, at 60 cM for somatic cell score, at 74 cM for herd life, and at 11 cM for female fertility. The 0.95 confidence intervals for quantitative trait loci locations were 20 cM for kilograms of fat, 27 cM for fertility, and 51 cM for somatic cell score. Two loci affecting fertility at opposite ends of the chromosome are apparently segregating in the population. A quantitative trait locus for fertility near the centromere was confirmed by application of the modified granddaughter design to a single family. Estimated frequency of the economically favorable allele in the Israeli Holstein cattle was less than 0.5. Significant genetic gain for fertility seems possible by marker-assisted selection.     

鄂尔多斯盆地东北缘府谷剖面中二叠统盒8段沉积相特征

鄂尔多斯盆地苏里格东部地区中二叠统盒8段为主力产气层,厘清其沉积相特征对指导该地区的天然气勘探具有重要意义。鄂尔多斯盆地东北缘府谷剖面与苏里格东部地区具有相似的物源体系与沉积环境,通过对府谷剖面进行实测,结合剖面岩石颜色、岩石学特征、微量元素特征、岩相特征等相标志分析,对盒8段沉积相进行准确划分并明确其沉积特征,可以弥补苏里格东部地区盒8段研究大部分局限于钻井、地震等方面,缺乏露头沉积相支撑的不足。结果表明:府谷剖面盒8段以灰绿、黄绿色粗—中粒岩屑砂岩为主,结构成熟度中等—较低; 微量元素V/(V+Ni)值表明盒8段沉积时期为氧化—弱还原过渡环境; 共划分出12种岩相,其中,块状砾岩相(Gm)、槽状交错层理砂岩相(St)、板状交错层理砂岩相(Sp)、平行层理砂岩相(Sh)为河流沉积中的典型岩相,不同岩相组合构成盒8段沉积体两种不同的垂向沉积序列; 盒8_下段—盒8²_上段为辫状河沉积,盒8¹_上段为曲流河沉积; 辫状河的垂向沉积序列由滞留沉积(Gm-Gst-Gsp)→心滩(Gst-Sq-Sp-St-Gm-St)→河道充填(Gm-St-Sh-M)→废弃河道(Sh-Fr-Mh-Sh-Fr-Mh)→越岸沉积(Sh-Fr-Mh-Fr-Mh)→泛滥平原(M-Mh-Mc-Mh-M)等6类微相构成了一个二元结构不明显的正粒序沉积序列; 曲流河的垂向沉积序列由滞留沉积(Gm)→边滩(Sp-St-Sm-Sp-St-Sh)→河漫滩(M-Mh-Mc(Fr)-Mh-M)等3类微相构成了一个典型二元结构沉积序列。     

花生矮化病毒病抗性SSR标记

利用抗、感矮化病毒病的花生品种ICGV86699和远杂9102为亲本配制杂交组合,构建重组近交系群体（RIL）,采用SSR技术和BSA分析方法,结合F6各个家系接种病毒后ELISA的鉴定结果,得到1个与花生矮化病毒病抗性连锁的分子标记XY38。此标记与抗性基因间的遗传距离为7.5cM,具有作为抗性育种辅助选择技术的潜力。     

Isolation of High Oleate Recombinants in Peanut by Near Infra‐Red Spectroscopy and Confirmation With Allele Specific Polymerase Chain Reaction Marker

Near infrared (NIR) spectrophotometer offers rapid, noninvasive, nondestructive, and high‐throughput phenotyping of seed samples for use in agriculture and industry. In this study, a reflectance‐based NIR spectrophotometer was calibrated and used for the isolation of desirable higher‐oleic‐acid peanut recombinants from single‐seed‐derived segregating populations at F₇ and F₈ generations. A calibration model was developed through partial least‐square regression using wet chemistry data from 158 peanut genotypes. Desirable prediction for oil, palmitic acid, oleic acid, and linoleic acid in intact seed was obtained based on this calibration. It detected significant high correlations (r) and coefficient of determination (R²) between the actual gas chromatography values and NIR predicted values of fatty acid profile in another 123 peanut genotypes that were generated from crosses involving a high‐oleate mutant and Spanish bunch varieties with early maturity. From this recombinant single‐seed‐derived progenies, 15 higher‐oleate recombinants were isolated and later genotyped through an in‐house developed polymerase chain reaction‐based allele specific marker. The present study has generated high‐oleate peanut recombinants with early maturity in Spanish bunch background. The breeding materials generated here will be evaluated for yield attributing traits at different locations in future.     

地下燃气管道电子标识及智能化管理系统研究

分析了北京地下燃气管网在实际运行维护中出现的问题,论述了地下燃气管道电子标识及智能化管理系统的组成、流程、功能以及该系统在实际工程中的实施情况。