Extension of cell membrane boosting squalene production in the engineered Escherichia coli

Squalene is a lipophilic and non-volatile triterpene with many industrial applications for food, pharmaceuticals, and cosmetics. Metabolic engineering focused on optimization of the production pathway suffer from little success in improving titers because of a limited space of the cell membrane accommodating the lipophilic product. Extension of cell membrane would be a promising approach to overcome the storage limitation for successful production of squalene. In this study, Escherichia coli was engineered for squalene production by overexpression of some membrane proteins. The highest production of 612 mg/L was observed in the engineered E. coli with overexpression of Tsr, a serine chemoreceptor protein, which induced invagination of inner membrane to form multilayered structure. It was also observed an increase in unsaturated fatty acid in membrane lipids composition, suggesting cellular response to maintain membrane fluidity against squalene accumulation in the engineered strain. This study potentiates the capability of E. coli for squalene production and provides an effective strategy for the enhanced production of such compounds.     

从抗体阳性的甲型肝炎接触者粪便中分离出一株甲型肝炎病毒

几年来,国内外已相继用组织培养法分离多株甲型肝炎病毒(HAV),并已证实了甲型肝炎患者及亚临床型感染在传染本病中的重要性。但HAV在流行点的正常人群中存在短期携带的问题还尚未见报道。本文采用人胚肺二倍体细胞(2BS)从甲型肝炎接触者粪便中分离出一株HAV,并经免疫电镜、免疫荧光及参比血清鉴定等证实。现将结果报告如下。     

长春和北京地区引起婴幼儿肺炎的3型与7型腺病毒的分子流行病学研究

对长春和北京地区连续12年(1976年冬至1988年春)引起小儿肺炎的3、7型腺病毒102株标本,进行了限制性内切酶核酸电泳图谱分析。56株7型腺病毒经BamHⅠ、BclⅠ、BglⅠ、XbaⅠ、SmaⅠ、HindⅢ分析后,表现为两个基因组型——Ad7 b和Ad7 d。46株3型腺病毒被Bg1 Ⅱ、BamHⅠ酶解后,表现为 3个基因组型——Ad 3Ⅰ、Ad 3Ⅱ、Ad 3Ⅲ。各基因组型的分布情况是:56株7型腺病毒中,43株为Ad 7 b(76.8％),流行于1976年冬至1986年春;13株是Ad 7 d(23.2％),出现于1982年,与Ad 7 b共同流行;1986年～1988年分析的5株病毒都是Ad 7d。43株3型腺病毒中,Ad3Ⅰ42株(91.0％),分布于12年中;Ad 3Ⅱ、Ad 3Ⅲ各2株,散在分布。此结果表明,国内这12年中引起小儿肺炎的3型腺病毒至少有3个基因组型,7型腺病毒至少有两个基因组型。Ad3Ⅰ和Ad7 b是流行优势基因组型。但自80年代初开始出现Ad7 d以来,有逐年增多的趋势,最近两年的标本又都是Ad7 d,很可能它将取代Ad7 b而成为流行的优势基因组型.     

粗皮松萝群体分化性状的研究

本文对于长白山南坡高山半寒漠植被中大面积匍匐生长在地表的粗皮松萝群体从形态学、生态学、化学及分类学方面进行了初步研究。在分类学上将这一地生群体命名为匍匐变种;在种群落结构方面进行了样方调查;从生物系统学角度进行了初步分析。     

抗白粉病小麦染色体组型的分子标记与生化标记分析

应用与小麦第六同源群有关的分子和生化标记,包括ＤＮＡ探针ｐＳｃ５·３Ｈ３和ｐＳＲ１６７以及同工酶Ｅｓｔ－５和ａ－Ａｍｙ－１,对来自六倍体小黑麦Ｂｅａｇｌｅ与普通小麦科冬５８杂交后代Ｆ１花粉植株的抗白粉病株系Ｍ２４．Ｍ０９及Ｍ１７进行了分析。结果表明,Ｍ２４、Ｍ０９及Ｍ１７不同程度地含有黑麦染色体成分,而且电泳谱带差别较大,据此推断,Ｍ０９为６ＲＬ的易位系。因此,生化和分子标记不仅可以用于确定外源片段的存在,而且可以帮助确定染色体组型和外源片段的位置     

Galectin‐3 mediates pulmonary vascular endothelial cell dynamics via TRPC1/4 under acute hypoxia

Galectin‐3 (Gal‐3) has been implicated in various biological functions, yet little is known about its role in regulating the dynamics of pulmonary vascular endothelial cells. Gal‐3 was shown to be increased in hypoxic model rats by sequencing analysis. We exposed pulmonary vessel endothelial cells (PVECs) to hypoxia or Gal‐3 stimulation, following which cell apoptosis and autophagy were measured with the relevant methods. The results demonstrated that hypoxia elevated nuclear factor‐κB (NF‐κB) activity and Gal‐3 expression. Gla‐3 decreased the expression of Bcl‐2, Alix, Beclin‐1, Atg5, and LC3A/B. The messenger RNA and protein levels of transient receptor potential channel 1/4 (TRPC1/4) and calpain were reduced after Gal‐3 treatment. Gal‐3 also activated protein kinase B/glycogen synthase kinase‐3 β/mammalian target of rapamycin signaling pathways in PVECs. These results suggest that a hypoxia‐mediated increase in Gal‐3 promotes apoptosis and inhibits autophagy by inhibiting the TRPC1/4 pathway and activating the protein kinase B/glycogen synthase kinase‐3 β/mammalian target of rapamycin signaling pathway in PVECs. Furthermore, these results may provide us with a new direction to explore the pathogenesis of pulmonary artery hypertension.     

温度诱导Targetron系统用于大肠杆菌高效基因失活

构建基于TeI3c/4c嗜热二型内含子的温度诱导Targetron基因失活系统 (Thermotargetron),并应用于中温微生物基因编辑。在大肠杆菌HMS174 (DE3) 基因组中,选择Subunit of flagellum基因 (fliC) 和C4 dicarboxylate orotate:H+ symporter基因 (dctA) 为靶基因。根据TeI3c/4c DNA识别规则,在fliC和dctA基因中选择fliC489a、fliC828s、fliC1038s和dctA2a位点为基因打靶位点。使用重叠延伸PCR方法,基于pHK-TT1A质粒构建打靶载体。打靶载体转化HMS174菌株,对数期转化子培养液48 ℃热激1 h后涂布于氯霉素抗性LB平板上。使用菌落PCR和DNA测序检测突变株并计算基因失活效率。获得突变株后,通过琼脂穿刺和碳源代谢实验,鉴定ΔfliC、ΔdctA突变株表型变化。菌落PCR测序结果表明,TeI3c/4c插入到fliC和dctA基因设计位点,且打靶效率高达100%。突变株表型验证实验表明,ΔfliC突变株运动能力显著下降,ΔdctA突变株苹果酸代谢能力缺失。综上所述,文中建立了一套适用于嗜中温微生物的温度诱导型、高效基因失活系统,该系统可通过控制宿主菌在48 ℃保温时间实现高效、靶向、精准基因失活。     

围封对天山高寒草原4种植物叶片和土壤化学计量学特征的影响

以中国科学院巴音布鲁克草原生态系统研究站长期围栏内外的羊茅(Festuca ovina)、天山赖草(Leymus tiansecalinus)、二裂委陵菜(Potentilla bifurca)和鹅绒委陵菜(Potentilla anserine)4种植物叶片和土壤为研究对象,分析了放牧与围封对植物叶片和土壤C、N、P的化学计量特征的影响。结果表明,围封样地土壤养分浓度整体高于放牧样地(P0.05),全氮(TN)浓度除外。围封显著增加羊茅叶片C、N浓度(P0.05),对P浓度影响不显著;围封显著增加鹅绒委陵菜叶片的C浓度,但是显著降低叶片的N和P浓度(P0.05),围封对天山赖草和二裂委陵菜养分含量影响不显著。围封显著增加鹅绒委陵菜C∶N和C∶P(P0.05);围封显著降低羊茅C∶N、C∶P和增加N∶P(P0.05);围封显著降低二裂委陵菜C∶N(P0.05),对天山赖草的化学计量特征影响不显著。不同植物对围封的响应不同,意味着长期围封可能会改变天山高寒草原生态系统的结构。围封降低优势种(羊茅)的固碳能力,增加退化期出现的代表性植物(鹅绒委陵菜)的固碳能力,表明在长期围封下植物凋落物中的杂类草(鹅绒委陵菜)可能更多的为土壤提供碳来源,也能促进优势禾本科物种的氮含量和碳含量的增加。