Generation of formic acid and ethanolamine from serine in biosynthesis of linear gramicidin by a cell-free preparation of Bacillusbrevis (ATCC 8185)

A growing organism that produces antibiotic peptide was incubated with L-(U-¹⁴C)serine for labeling linear gramicidin. Linear gramicidin was isolated by a simple chromatographic method from tyrothricin (mixture of linear gramicidin and tyrocidine) applied to a column of basic aluminum oxide. The hydrolysate of labeled linear gramicidin on thin layer chromatography showed that L-(U-¹⁴C)serine was one of a precursor of ethanolamine moiety by autoradiography. L-(3-¹⁴C)serine generated formic acid in the presence of tetrahydrofolic acid by an enzyme fraction prepared with ammonium sulfate, and further formed ethanolamine binding to the protein. Formylvaline was biosynthesized by it with tetrahydrofolic acid and ATP, and subsequently released from the protein.     

Maturation and fertilization of porcine oocytes in vitro

Four experiments were conducted to study 1) factors affecting porcine oocyte maturation in culture medium and 2) a new method for oocyte maturation outside the porcine body. In Experiment 1, five groups of oocytes were cultured in m-TCM199 or m-KRB medium for 24 to 28, 32 to 36 or 40 to 42 hours and then were fertilized in vitro. The cleavage rate (two to four-cell stage) of oocytes cultured for 32 to 36 hours was significantly higher than those of the other oocytes. The results indicate that a suitable culture period for the in vitro maturation of porcine oocytes is 32 to 36 hours. In Experiment 2, four groups of oocytes were cultured in m-KRB or m-KRB supplemented with PFF, PMSG or FSH for in vitro maturation, and the cleavage rates of oocytes were 7.94, 22.56, 30.23 and 23.26%, respectively, after in vitro fertilization. The results show that porcine follicular fluid (PFF) and gonadotrophins added to the culture medium promote porcine oocyte maturation in vitro. In Experiment 3, oocytes were cultured in m-KRB or m-TCM199, supplemented with both gonadotrophin and pocine folliclar fluid for maturation in vitro. After fertilization in vitro, the cleavage rates of oocytes were 26.32 and 27.93% for the two media. The results indicate that the difference between m-KRB and m-TCM199 was insignificant when the media were used to culture porcine oocytes. But there was a significant difference when PFF and gonadotrophins were added to the basic media. In Experiment 4, porcine oocytes were transferred into the reproductive tracts of other animals for maturation. After 34 to 36 hours, the oocytes were collected and fertilized in vitro. The cleavage rates of oocytes were 10.42, 28.45, 3.33 and 36.36%, respectively, for the oocytes matured in mouse uterine horns, rat uterine horns, rat oviducts or rabbit oviducts. The results show that porcine oocytes can be matured in the reproductive tracts of other animals.     

基于MaxEnt模型的甘肃安西极旱荒漠国家级自然保护区北山羊生境评估

明晰甘肃安西极旱荒漠国家级自然保护区珍稀濒危物种北山羊的分布格局,并阐释气候变化和人类活动对北山羊的影响,对今后北山羊生境管理和物种保护具有重要意义。基于北山羊实测分布点记录和环境变量数据,结合MaxEnt模型和ArcGIS空间分析功能,利用CMIP6的8个气候模式均值预测中度发展路径（SSP2-4.5）下,基准期（1970-2000年）和未来气候（2041-2060年、2081-2100年）变化情景下,甘肃安西极旱荒漠国家级自然保护区北山羊的潜在适生区分布范围及变化,并综合贡献率和置换重要性值对北山羊生境选择关键环境因子进行了分析。研究结果表明：（1） MaxEnt模型的预测精度较高,三种气候条件下ROC曲线下面积（AUC,Area Under Curve）>0.97,且真实技巧统计（TSS,True Skill Statistic）>0.90,模拟结果可靠。（2）影响北山羊生境选择的主要环境因子为气候条件（降水量季节性变异系数和等温性）、海拔和人为干扰（距泉和居名点距离）。水是保护区北山羊生存的最基本要素,气候条件共同控制北山羊生境条件。此外,北山羊习性决定其生境宜选择高海拔和远离人类活动影响地区。（3）基准期保护区北山羊主要分布在北片和南片高海拔山区,面积365.77 km²（占整个保护区的4.31%）,北山羊适生区面积北片>南片、中高等适生区主要位于保护区北片。（4） CMIP6未来气候变化情景下,随着保护区生态环境改善和濒危物种保护措施的实施,北山羊潜在适生区面积呈增加趋势,但是受北山羊近亲繁殖的影响,整体上北山羊数量和适生区面积增加并不显著且有向南部及高海拔地区转移趋势。     

城市“源-汇”热景观变化及其空间作用强度特征——以深圳西部地区为例

城市热岛作为一种整体涌现性的地球表层系统环境问题受到地理学和景观生态学的广泛关注,基于空间异质性对城市冷热岛影响方面的研究初步揭示了城市热环境的形成机制,然而城市冷热岛在空间上是否存在相互影响及相互作用强度还有待研究。以深圳市西部城市区为例,应用地表温度和景观分类数据,通过构建二维矩阵识别"源-汇"热景观,并采用"源-汇"热贡献指数和空间引力模型等方法定量分析了城市"源-汇"热景观的演变过程,"源-汇"热贡献程度的时空分异特征,以及两者在空间上的整体相互作用强度及演变特征。结果表明：（1）1988-2019年,城市热源、汇在空间上呈现由包围到反包围的变化,热源景观呈现基质化,其面积增长210.45%;热汇景观则表现为破碎化趋势,其面积减少48.07%。与之对应的热汇景观地表温度与区域的平均温度的温差逐渐增大,从1988年的0.49℃上升到2019年的1.91℃。（2）源、汇对热环境的贡献规模呈增长趋势,热源景观的贡献程度会随着热岛规模从零星状向区域化的转变过程中提升,形成温度聚集效果;而热汇景观的贡献程度会随着冷岛规模的破碎化过程中被抑制。（3）1988-2019年"源-汇"热景观的作用对基本保持稳定,但总作用强度发生了显著的变化,30年间增长了8倍。从热汇景观一对多影响热源景观转变为热源景观一对多影响热汇景观,且作用强度也越来越强。研究发现城市热"源-汇"的空间相互作用强度与"源-汇"热景观的贡献程度有密切关联,因此控制热源景观规模和维护热汇景观规模对调控城市热环境,对提升城市居民生活质量有重要意义。     

浙江西门岛海洋特别保护区大型底栖动物功能群特征及其与环境的关系

为研究浙江西门岛海洋特别保护区大型底栖动物功能群的变化规律及其与环境因子的关系,作者分别于2010年4月(春季)、11月(秋季),2011年8月(夏季)和2012年2月(冬季)进行了4个航次的大型底栖动物调查,共鉴定出大型底栖动物78种,根据其食性类型划分为浮游生物食者、植食者、肉食者、杂食者、碎屑食者5种功能群.各功能群平均密度从高到低依次为浮游生物食者＞肉食者＞植食者＞碎屑食者＞杂食者,平均生物量从高到低依次为浮游生物食者＞碎屑食者＞肉食者＞杂食者＞植食者.单因素方差分析结果表明,大型底栖动物各功能群的密度和生物量季节间均无显著性差异.典范对应分析结果表明,影响大型底栖动物功能群的主要环境因子包括温度、溶解氧、溶解态无机磷和表层沉积物的中值粒径,排序轴对功能群-环境关系的贡献率计算结果表明环境变量可以较好地解释功能群的变化情况.     

Systematic Evaluation of Protein Sequence Filtering Algorithms for Proteoform Identification Using Top‐Down Mass Spectrometry

Complex proteoforms contain various primary structural alterations resulting from variations in genes, RNA, and proteins. Top‐down mass spectrometry is commonly used for analyzing complex proteoforms because it provides whole sequence information of the proteoforms. Proteoform identification by top‐down mass spectral database search is a challenging computational problem because the types and/or locations of some alterations in target proteoforms are in general unknown. Although spectral alignment and mass graph alignment algorithms have been proposed for identifying proteoforms with unknown alterations, they are extremely slow to align millions of spectra against tens of thousands of protein sequences in high throughput proteome level analyses. Many software tools in this area combine efficient protein sequence filtering algorithms and spectral alignment algorithms to speed up database search. As a result, the performance of these tools heavily relies on the sensitivity and efficiency of their filtering algorithms. Here, we propose two efficient approximate spectrum‐based filtering algorithms for proteoform identification. We evaluated the performances of the proposed algorithms and four existing ones on simulated and real top‐down mass spectrometry data sets. Experiments showed that the proposed algorithms outperformed the existing ones for complex proteoform identification. In addition, combining the proposed filtering algorithms and mass graph alignment algorithms identified many proteoforms missed by ProSightPC in proteome‐level proteoform analyses.     

基于同源重组的裂殖壶菌遗传转化体系的构建及验证

【目的】裂殖壶菌是一种能高效生产DHA的海洋真菌;基因工程技术已经成功应用在微生物改造和代谢机理研究中,利用基因工程技术对裂殖壶菌进行改造首先需要构建适合裂殖壶菌的遗传转化体系;【方法】本文利用电转化的方法将含有18S r DNA同源重组片段的ble基因导入裂殖壶菌中,通过zeocin抗性平板筛选出阳性菌株,并设计ble基因引物,以裂殖壶菌基因组为模板,进行PCR验证ble基因是否成功结合到裂殖壶菌染色体上。【结果】筛选获得的抗性菌株基因组上确实PCR出ble基因片段,对改造菌株与原始菌株进行发酵培养,发现改造后菌株在生物量、油脂含量、DHA含量及脂肪酸分布等方面和原始菌株基本一致。【结论】抗性基因的插入不会影响菌株的正常代谢,该体系的构建为后续其他外源基因导入奠定基础。     

高山被孢霉产花生四烯酸及其遗传改造的研究进展

花生四烯酸作为一种重要的多价不饱和脂肪酸,因其具有多种生理功能而被认为是潜在的食品添加剂和药物。近年来,利用高山被孢霉合成花生四烯酸已成为研究热点。前期相关研究主要集中在菌种选育及发酵调控方面。随着研究的不断深入,关于高山被孢霉合成花生四烯酸的代谢途径的研究取得了较大进展。以下简要概述前期工作进展,着重论述花生四烯酸合成途径的关键酶及其高山被孢霉的遗传改造的研究情况,包括生物合成花生四烯酸代谢途径、关键酶及其应用、高山被孢霉的遗传操作系统的构建以及遗传改造的应用,并对其研究前景进行了展望。     

RETRACTED ARTICLE: Therapeutic DNA Vaccination as a Repair Strategy Following Spinal Cord Injury

Myelin-derived proteins, such as tenascin-R (TN-R), myelin associate glycoprotein (MAG), oligodendrocyte-myelin glycoprotein (OMgp), and Nogo-A, inhibit the central nervous system regeneration. In this study, the DNA vaccine encoding for oligodendrocyte and myelin-related antigens was employed to attenuate the axonal growth inhibitory properties of myelin in the setting of spinal cord injury. Using a rat spinal cord dorsal hemisection model, the vaccine directed against the inhibitory epitopes of Nogo-A, MAG, OMgp, and TN-R was administered intramuscularly once a week following spinal cord injury, supplemented with local application of specific anti-sera against the four antigens. Anterograde labeling of dorsal column fibers showed active axonal regeneration through the lesion site at the eighth week following the treatment in experimental group but not in control groups. Light microscopic and ultrastructural analysis revealed that vaccination with these myelin-related antigens did not lead to demyelinating disease. OMgp and TN-R levels were down-regulated at the lesion site together with a parallel increase in growth-associated protein 43 levels in the treatment groups. This study reveals the effective approach of a DNA vaccine strategy by attaining the special antibody to direct neutralization of the myelin inhibitors during spinal cord injury.