氨基酸介导的TOR信号传导通路研究进展

TOR(target of rapamycin)是一种进化上十分保守的丝氨酸(Ser)/苏氨酸(Thr)蛋白激酶,可以感知营养状况、能量、生长因子等信号,进而调节细胞的生长、增殖和凋亡等生理进程.本文综述了TOR的上、下游信号传导通路及各种氨基酸对TOR信号传导通路的影响,为探讨氨基酸调控蛋白质合成的作用机制及建立精确...     

雷帕霉素预处理延缓牡丹‘洛阳红’切花衰老的生理效应

为研究雷帕霉素靶标激酶TOR在牡丹开花衰老过程中的生理作用，以牡丹（Paeonia suffruticosa Andr.）‘洛阳红’切花为试材，用0.01 μmol • L-1雷帕霉素预处理2 h后瓶插，并测定瓶插寿命、能量物质含量、糖分含量、乙烯释放速率、呼吸速率、MDA含量以及PsTOR、PsSnRK1和PsHXK1表达量变化。结果表明：雷帕霉素预处理可延长该牡丹切花的瓶插最佳观赏期和增大花朵的最大花径；另外，雷帕霉素预处理还可提高花瓣可溶性糖含量和能荷积累，降低呼吸耗能和MDA含量，并下调瓶插初期PsSnRK1和PsHXK1的表达水平，上调瓶插后期PsTOR和PsSnRK1的表达水平。说明雷帕霉素可通过TOR途径调控能量感知，延缓牡丹切花的开放和衰老进程，进而提高瓶插品质。     

营养介导的TOR信号传导研究进展

TOR(target of rapamycin)是一种结构和功能高度保守的蛋白激酶,它能够通过整合营养、能量、生长因子等信号调节细胞的增殖、体积增大及细胞周期进程。作者综述了TOR激酶的上下游信号传导途径及氨基酸和能量等营养状况对这些信号传导通路和细胞生长的影响,为探讨营养调控体蛋白合成的机制及精准营养供给技术的建立提供宝贵的理论基础和开创性思路。     

Optimal dietary alpha‐linolenic acid/linoleic acid ratio improved digestive and absorptive capacities and target of rapamycin gene expression of juvenile grass carp (Ctenopharyngodon idellus)

Growth performance, digestive and absorptive capacities and target of rapamycin (TOR), ribosomal protein S6 kinase 1 (S6K1) and eIF4E‐binding protein (4E‐BP) gene expression in the hepatopancreas and intestine of juvenile grass carp (Ctenopharyngodon idellus) fed graded ratios of dietary alpha‐linolenic acid/linoleic acid (ALA/LNA) (0.01, 0.34, 0.68, 1.03, 1.41, 1.76 and 2.15) for 60 days were investigated. The results showed that ALA/LNA ratio of 1.03 significantly improved (i) per cent weight gain (PWG) and feed efficiency, (ii) hepatopancreatic trypsin, chymotrypsin, lipase, amylase and intestinal creatine kinase (CK) activities, (iii) hepatopancreatic trypsinogen‐2 and chymotrypsinogen mRNA levels. Meanwhile, fish fed with ALA/LNA ratio of 0.68 significantly enhanced, (iv) Na⁺/K⁺‐ATPase and γ‐glutamyl transpeptidase activities in whole intestine, and alkaline phosphatase activities in the proximal intestine (PI) and distal intestine, (v) amylase, intestinal Na⁺/K⁺‐ATPase alpha‐subunit isoform 1, Na⁺/K⁺‐ATPase alpha‐subunit isoform 8 and CK mRNA abundances, (vi) TOR and S6K1 gene expression in the hepatopancreas and intestine of juvenile grass carp. Based on the quadratic regression analysis of PWG, cholecystokinin and leptin contents in the PI, optimal dietary ALA/LNA ratio of juvenile grass carp (8.78–72.00 g) was estimated to be 1.08, 1.19 and 1.05, respectively.     

二穗短柄草BdTOR基因的克隆及功能分析

在酵母、植物、动物和人类中,TOR是一个进化保守的重要调节因子,它通过整合营养和能量信号促进细胞增殖和生长。从二穗短柄草中克隆得到全长7 401 bp的Bd TOR基因,编码2 466个氨基酸残基,预测其具有HEAT重复域、FAT域、FRB域、激酶域、FATC域等保守结构域。实时定量PCR表明Bd TOR在二穗短柄草组织器官中相对表达量由高到低依次为幼穗幼苗根叶茎;Bd TOR及其激酶域的瞬时表达表明其定位于拟南芥原生质体的细胞核和细胞质,并在细胞质中区域化分布;Bd TOR具有促进根、叶生长,增加生物量,提高生长速率的功能;过表达Bd TOR可以回补雷帕霉素对Sc FKBP12过表达株系BP12的抑制;在雷帕霉素处理下Bd TOR能够与Sc FKBP12互作。     

营养与鱼类免疫研究进展

鱼类生长发育与疾病抵抗力密切相关,而营养物质在鱼类疾病抵抗力及其免疫调控中发挥着重要作用.已有研究表明部分营养物质可通过雷帕霉素靶蛋白(TOR)和核因子E2相关因子2(Nrf2)信号途径调控鱼类免疫器官细胞因子产生,降低鱼类免疫器官氧化损伤,从而保证其结构完整性和功能正常,进而提高鱼类非特异性和特异性免疫力,增强鱼类疾病抵抗力.本文就部分氨基酸(蛋氨酸和异亮氨酸)、脂溶性维生素(维生素A)、水溶性维生素(维生素B1、吡哆醇、泛酸、胆碱和肌醇)以及营养性添加物(蛋氨酸羟基类似物)等营养物质与鱼类疾病抵抗力、非特异和特异性免疫防御、免疫器官生长发育、结构完整性和抗氧化防御、细胞因子及其相关信号途径之间关系的研究进展作一综述.     

绿盲蝽雷帕霉素靶蛋白的克隆、抗体制备及在蜕皮激素诱导下的应答

【目的】克隆绿盲蝽（Apolygus lucorum）雷帕霉素靶标全长基因（AlTOR）,研究AlTOR时空表达特性及在外源蜕皮激素（20E）诱导下的应答响应,进一步获得原核表达的重组蛋白及制备多克隆抗体,分析该抗体的特异性,为后续研究AlTOR蛋白功能打下基础。【方法】RACE法克隆AlTOR基因序列全长,qRT-PCR分析AlTOR表达特性以及在20E及其抑制剂U73122诱导下的应答响应;将含有AlTOR序列T载体经EcoR I及Xho I双酶切,构建原核表达载体pCzn1-AlTOR,经20℃、0.5 mmol·L^-1 IPTG诱导表达、变性、复性和蛋白纯化后,以获得AlTOR重组蛋白,最后再通过免疫,制备AlTOR蛋白多克隆抗体,Western blot评价该抗体的特异性。【结果】AlTOR的开放阅读框编码包含435个氨基酸,具有典型的TOR基因序列特征,即SIN1结构域、高度保守的CRIM结构域及PH域;AlTOR在绿盲蝽不同日龄、不同组织中均有表达,且在1日龄以及脂肪体中表达量最高;20E可诱导绿盲蝽不同日龄若虫期AlTOR的表达,同时也能诱导成虫头、翅、卵巢和脂肪体中AlTOR表达上调,分别上调200.00%、118.89%、20.53%和60.82%,而U73122在中肠和卵巢中会显著抑制AlTOR的表达。经双酶切的重组克隆载体可成功亚克隆到pCzn1载体上,命名为pCzn1-AlTOR;IPTG诱导的重组质粒可特异性表达一个约36 kD的蛋白,且主要以包涵体形式存在;经Ni-IDA亲和层析纯化后,仅在36 kD附近有一条明显的特异性条带。进一步免疫及间接ELISA方法确定抗血清对TOR蛋白的效价,Western blot分析表明制备的多克隆抗体可以特异性与AlTOR重组蛋白及绿盲蝽3龄若虫总蛋白结合,说明制备的AlTOR多克隆抗体特异性较好,可以用于后续的蛋白研究。【结论】AlTOR在绿盲蝽体内的表达谱显示出发育阶段特异性和组织特异性;20E及其抑制剂诱导后,AlTOR呈现出相反的应答反应;本研究获得的AlTOR重组蛋白的多克隆抗体具有高特异性。     

Effects of breed and dietary nutrient density on the growth performance,blood metabolite,and genes expression of target of rapamycin (TOR) signalling pathway of female broiler chickens

This study was conducted to compare the effects of exchanged diets with identical energy level on characteristics of slow‐growing (WENs Yellow‐Feathered Chicken, WYFC) and fast‐growing (White Recessive Rock Chicken, WRRC) female chickens. A total of 1450 WYFC and 1150 WRRC 1‐day‐old female hatchlings were used. A high‐nutrient‐density (HND) diet and a low‐nutrient‐density (LND) diet were formulated for three phases. A completely randomized experimental design with a 2 × 2 factorial arrangement (diet and breed), each with five replicates of 145 and 115 birds, was applied. The results showed that WRRC had a higher body weight (BW), average daily feed intake and average daily gain than WYFC throughout the experiment (p < 0.05). WYFC that were provided with HND groups had a higher BW only in the starter and grower phases, whereas WRRC had a higher BW in the HND group than in LND groups throughout the experiment. The feed:gain ratio and protein efficiency ratio (PER) were better for WRRC in the starter and grower phases; however, these ratios were better for WYFC in the finisher period. The LND groups had a higher PER throughout the experiment for both breeds (p < 0.05). The breast and leg muscle weights were higher for WRRC compared with WYFC during the grower and finisher phases (p < 0.05). WRRC had a lower liver index but higher serum UA and alkaline phosphatase (ALP) concentrations than WYFC (p < 0.05). No diet effect was observed on organ indices, muscle yields or blood responses. The gene expressions of Rheb, TOR, S6K1 and 4E‐BP1 in gastrocnemius muscle were the highest in the WYFC‐LND groups at 63 and 105 days (p < 0.05). These findings suggested that different genotypes respond differently to changes in dietary nutrient density and that lower‐nutrient‐density diets are optimal for the long‐term housing of broiler chickens.     

三次采油用速溶型超高分子量聚丙烯酰胺的合成

为满足三次采油要求,通过丙烯酰胺(AM)溶液聚合制备了分子量大于1.7×107,溶解时间小于30min,过滤比小于2的速溶型超高分子量聚丙烯酰胺.其最佳合成条件为:AM30%;过硫酸铵0.02%;三乙胺0.02%;引发温度10～15℃;EDTA-2Na 0.08%;尿素3%;甲酸钠0.2%.探讨了聚合工艺条件和聚合配方对聚合物增粘性能和溶解性能的影响规律.