牛α干扰素在毕赤酵母中的高效分泌表达及活性鉴定     

王延群  张璐  李玉明  汪志艳  金家民  刘永刚  王刚  王淑杰  姜成刚 《中国预防兽医学报》2013,35(10)

为高效分泌表达牛α干扰素(boIFN-α),本研究通过人工合成boIFN-α基因,将目的基因克隆至表达载体pPIC9K中,构建重组质粒pPIC9K-boIFN-α,将其电转化于毕赤酵母菌株GS115,利用抗药选择压力G418筛选重组菌,对重组菌诱导表达,取上清进行SDS-PAGE和western blot检测,并优化重组菌的诱导表达条件.结果显示:筛选获得高效分泌表达boIFN-α的重组菌,其最佳诱导条件为:250 r/min,26℃培养,1％甲醇浓度诱导,诱导72 h.上清中目的蛋白表达量最高可达200 μg/mL,本研究为boIFN-α在生产中的应用奠定了基础.  相似文献   

Increased Inflammatory Mediators in Horses Naturally Infected with Trypanosoma vivax. A Preliminary Study     

Aleksandro S. Da Silva  Marta M.M.F. Duarte  Guilherme V. Bochi  Rafael N. Moresco  Lenita Moura Stefani  Silvia G. Monteiro 《Journal of Equine Veterinary Science》2013

The aim of this study was to measure the levels of inflammatory mediators in serum from horses naturally infected with Trypanosoma vivax. Banked serum samples collected during a previously reported T. vivax natural infection were used to analyze proinflammatory cytokines such as interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NO_x) levels. We evaluated 12 serum samples from horses from a farm in southern Brazil, four of which had parasitological and molecular diagnoses for T. vivax and presented with clinical signs of disease. Cytokines were assessed by quantitative sandwich enzyme-linked immunosorbent assay, and NO_x was measured using the modified Griess method. Levels of IFN-γ, TNF-α, IL-1, IL-6, and NO_x were increased in serum of infected animals compared to that in noninfected animals. Therefore, infection with T. vivax caused an increase in proinflammatory cytokines and nitric oxide content.  相似文献   

Role of estrogen receptor-α (ESR1) and the type 1 insulin-like growth factor receptor (IGFR1) in estradiol-stimulated proliferation of cultured bovine satellite cells     

E. Kamanga-Sollo  M.E. WhiteW.J. Weber  W.R. Dayton 《Domestic animal endocrinology》2013

Although the exact mechanism(s) by which estradiol (E₂) enhances muscle growth in a number of species, including humans and cattle, is not known, E₂ treatment has been shown to stimulate proliferation of cultured bovine satellite cells (BSCs). This is particularly significant because satellite cells are the source of nuclei needed to support postnatal muscle fiber hypertrophy and are thus crucial in determining the rate and extent of muscle growth. The objective of this study was to assess the role of estrogen receptor-α (ESR1) and the type 1 insulin-like growth factor receptor (IGFR1) in E₂-stimulated proliferation of cultured BSCs. To accomplish this, we have used small interfering RNA (siRNA) to silence expression of ESR1 or IGFR1 and assessed the effects on E₂-stimulated proliferation in BSC cultures. In BSCs treated with nonspecific siRNA, E₂ significantly (P < 0.05) stimulates proliferation under conditions in which neither IGF-1 nor IGF-2 expression is increased; however, treatment of ESR1- or IGFR1-silenced cells with E₂ does not significantly stimulate proliferation. These results indicate that both ESR1 and IGFR1 are required for E₂ to stimulate proliferation in BSC cultures. The fact that this occurs under culture conditions in which neither IGF-1 nor IGF-2 mRNA expression is increased strongly suggests that E₂ activates IGFR1 via a mechanism that does not involve increased IGF-1 or IGF-2 binding to the receptor.  相似文献   

Chlamydia infection in cats in New Zealand     

T.J. Gruffydd-Jones  B.R. Jones  H. Hodge  M. Rice  M.A. Gething 《New Zealand veterinary journal》2013,61(5):201-203

Conjunctival swabs collected in 1991-92 from 333 pedigree and non-pedigree cats were tested for the presence of Chlamydia spp. antigen using an ELISA antigen kit. Forty (18.4%) of the 217 samples from cats with conjunctivitis were positive. Seven (6%) of 116 samples from cats which were in contact with cats with conjunctivitis but which showed no clinical signs at the time of sample collection were positive. Positive-testing cats were frequently from multi-cat households. Chlamydia spp. is present and associated with conjunctivitis in cats in New Zealand. Infection may occur concurrently with viral diseases. Feline calicivirus was recovered from 27 (21 with conjunctivitis) of 37 cats tested in five catteries. Four cats (with conjunctivitis) were FIV-positive.  相似文献   

Pregnancy recognition signaling mechanisms in ruminants and pigs     

Fuller  W  Bazer 《畜牧与生物技术杂志(英文版)》2013,(3):170-179

Maternal recognition of pregnancy refers to the requirement for the conceptus（embryo and its associated extraembryonic membranes） to produce a hormone that acts on the uterus and/or corpus luteum（CL） to ensure maintenance of a functional CL for production of progesterone;the hormone required for pregnancy in most mammals.The pregnancy recognition signal in primates is chorionic gonadotrophin which acts directly on the CL via luteinizing hormone receptors to ensure maintenance of functional CL during pregnancy.In ruminants,interferon tau（IFNT） is the pregnancy recognition signal.IFNT is secreted during the peri-implantation period of pregnancy and acts on uterine epithelia to silence expression of estrogen receptor alpha and oxytocin receptor which abrogates the oxytocin-dependent release of luteolytic pulses of prostaglandin F2-alpha（PGF） by uterine epithelia;therefore,the CL continues to produce progesterone required for pregnancy.Pig conceptuses secrete interferon delta and interferon gamma during the peri-implantation period of pregnancy,but there is no evidence that they are involved in pregnancy recognition signaling.Rather,pig conceptuses secrete abundant amounts of estrogens between Days 11 to 15 of pregnancy required for maternal recognition of pregnancy.Estrogen,likely in concert with prolactin,prevents secretion of PGF into the uterine venous drainage（endocrine secretion）,but maintains secretion of PGF into the uterine lumen（exocrine secretion） where it is metabolized to a form that is not luteolytic.Since PGF is sequestered within the uterine lumen and unavailable to induce luteolysis,functional CL are maintained for production of progesterone.In addition to effects of chorionic gonadotrophin,IFNT and estrogens to signal pregnancy recognition,these hormones act on uterine epithelia to enhance expression of genes critical for growth and development of the conceptus.  相似文献   

双酶法分解怀山药的工艺研究     

刘瑞山  梁治军  常路路  王新房  齐强强 《安徽农业科学》2013,(14):6462-6465

[目的]研究高温α-淀粉酶和糖化酶对怀山药的分解效果。[方法]利用高温α-淀粉酶和糖化酶对怀山药分步进行水解糖化以减小分散颗粒直径和沉淀率,在控制一定温度和时间的条件下,采用正交设计和因素分析来确定最适酶解条件,从而提高山药汁的可溶性固形物百分含量,降低其沉淀率和色泽。[结果]试验表明,双酶法分解怀山药的最适酶解条件为:高温α-淀粉酶添加量0.4%(20 000U),山药浓度10%,酶解温度85℃,时间2 h条件下酶解效果最佳;糖化酶添加量0.4%(10 000 U),糖化温度55℃,时间3 h,pH 5.0。[结论]研究可为怀山药的深度开发提供一定的理论依据。  相似文献   

水稻α-淀粉酶基因的表达模式与颖花开放的关系     

张绩  周上铃  何发  刘莉莎  张玉娟  何晋宇  杜晓秋 《中国农业科学》2023,56(7):1275-1282

【目的】淀粉降解与水稻浆片膨大和颖花开放过程密切相关,探究α-淀粉酶基因在颖花开放过程中的作用,为杂交水稻制种效率及产量的提高提供理论依据。【方法】在水稻扬花时,利用稀释碱性品红溶液进行离体穗子吸水试验,观察碱性品红在颖花中残留的组织,通过碘-碘化钾染色法确定11—14期（依据雄蕊发育分期）淀粉粒的分布变化,并通过RT-PCR、RT-qPCR和GUS报告基因检测多个α-淀粉酶基因在此期间的时空表达模式。【结果】水稻颖花开放前,内外稃片通过相互嵌合的钩合槽（marginal tissues of palea,mtp）将浆片和雌雄蕊封闭在内。当颖花开放时,浆片快速膨大,使得内外稃片的钩合点松开。扬花期间,离体穗子在稀释碱性品红溶液中吸水后,碱性品红染料主要残留在内外稃片钩合槽和浆片相连处组织以及花丝中。碘染试验显示,在12期（颖花开放前）,淀粉粒主要分布在雄蕊和内外稃片钩合槽,浆片中也有少量淀粉粒,在13—14期（颖花开放中）,内外稃片钩合槽和浆片中的淀粉粒均降解。RT-PCR分析发现OsRAmy2A和OsRAmy3D的表达量从12期开始上升,至13—14期表达量显著增强,到受精后1 d(...  相似文献   

大豆α-生育酚的遗传与QTL分析   总被引：1，自引：1，他引：0  

梁慧珍  余永亮  许兰杰  杨红旗  董薇  谭政伟  李磊  裴新涌  刘新梅 《中国农业科学》2019,52(1):11-20

【目的】 通过对大豆α-生育酚进行遗传和QTL分析,研究其遗传机制,定位其主效QTL,为高α-生育酚含量的大豆品种选育奠定遗传学基础。【方法】 以栽培大豆晋豆23为母本、山西农家品种大豆灰布支黑豆（ZDD02315）为父本杂交衍生的447个RIL作为供试群体构建遗传图谱,试验群体及亲本分别于2011年、2012年和2015年夏季在河南省农业科学院原阳试验基地种植,冬季在海南省三亚南繁基地种植。田间试验采取随机区组设计,2次重复。从6个环境中每个家系选取15.00 g籽粒饱满,大小一致的大豆种子,利用高效液相色谱法定性、定量测定样品中的α-生育酚含量。采用主基因+多基因混合遗传分离分析法和WinQTLCart 2.5复合区间作图法,对大豆α-生育酚含量进行主基因+多基因混合遗传分析和QTL定位。【结果】 基于主基因+多基因混合遗传分离分析法,α-生育酚受4对主基因控制,遗传基因分布在双亲中。4对主基因间加性效应值中3对为正值,表明这些基因来源于母本晋豆23;1对为负值,表明该对基因来源于父本灰布支黑豆;4对主基因之间相互作用的上位性效应表现为正值和负值的各有3对,说明不同基因间上位性效应对α-TOC的影响方向并不完全一致。环境因素引起的变异为0.13%—4.05%。表明α-TOC主要受4对主基因影响,受环境因素影响较小。采用WinQTLCart 2.5复合区间作图（CIM）共检测到17个影响α-生育酚的QTL,分布于第1、2、5、6、8、14、16、17共8条染色体中,单个QTL的贡献率8.35%—35.78%,QTL主要表现为加性效应。qα-D1a-1同时在2011年原阳、2012年原阳和三亚、2015年原阳4个环境下检测到,且均定位在第1染色体Satt320—Satt254标记区间19.79 cM处,解释的表型变异分别为12.55%、12.01%和11.89%、12.61%,加性效应值0.119-0.132,增加α-TOC含量的等位基因来自母本晋豆23;qα-A2-1同时在2011年原阳和三亚、2015年原阳3个环境下检测到,且均定位在第8染色体Sat_129—Satt377标记区间44.53 cM处,解释的表型变异分别为23.18%和22.56%、23.01%,加性效应值-0.195—-0.180,增加α-TOC含量的等位基因来自父本灰布支黑豆。qα-D1a-1和qα-A2-1 2个QTL能够稳定遗传。【结果】 α-生育酚最适遗传模型符合4MG-AI,即4对具有加性上位性效应的主基因遗传模型。其遗传主要受4对主基因影响,受环境因素影响较小。检测到α-生育酚的2个稳定主效QTL,Satt320—Satt254和Sat_129—Satt377是共位标记区间。  相似文献   

饲粮α-亚麻酸水平对意大利蜜蜂工蜂幼虫生理机能的影响     

于静  张卫星  马兰婷  胥保华 《中国农业科学》2019,52(13):2368-2378

【目的】探究饲粮中α-亚麻酸的添加水平对意大利蜜蜂（Apis mellifera ligustica）工蜂幼虫抗氧化活性和免疫能力的影响。【方法】移取1日龄意大利蜜蜂工蜂幼虫1 200只,随机分为5组,每组5个重复,每个重复48只;其中1组为对照组,饲喂不添加α-亚麻酸的基础饲粮,4组为处理组,分别饲喂α-亚麻酸添加水平为0.02%、0.04%、0.06%和0.08%的饲粮。按照室内蜜蜂幼虫饲养方法,将1日龄幼虫用移虫针移至温度适宜的加入200 μL饲粮的24孔细胞培养板内,培养板置于恒温培养箱中（温度33℃,相对湿度55%）,试验期间每天更换饲粮。饲养至第6天末或第7天初,幼虫开始有直立或排便现象时,将幼虫转移至提前铺好灭菌纸的24孔细胞培养板内准备化蛹。从饲养第1天开始,每天检查并记录幼虫和蛹的死亡数量,并将死亡个体及时移除,直至未死亡的蛹全部羽化新蜂,记录成功化蛹和羽化新蜂个体数量,统计幼虫化蛹率和羽化率。各组分别取5、6和7日龄幼虫测定抗氧化、免疫、脂质代谢指标及相关基因表达量。【结果】饲粮中α-亚麻酸的添加水平为0.02%和0.04%时,化蛹率和羽化率显著高于与其他处理组（P＜0.05）,而工蜂幼虫血淋巴中甘油三酯（TG）、总胆固醇（TC）和低密度脂蛋白（LDL）含量显著低于对照组,高密度脂蛋白（HDL）的含量却显著高于对照组（P＜0.05）。饲粮中α-亚麻酸的添加水平为0.04%时,工蜂幼虫超氧化物歧化酶（T-SOD）的活性较对照组显著增加,丙二醛（MDA）的含量显著降低（P＜0.05）。饲粮中α-亚麻酸的添加水平为0.02%、0.04%和0.06%时,6日龄工蜂幼虫的溶菌酶（lysozyme）和酚氧化酶（PO）活性显著高于对照组（P＜0.05）。饲粮中α-亚麻酸添加水平为0.04%时,6日龄工蜂幼虫脂肪酸合成酶（FAS）和乙酰辅酶A羧化酶（ACC）活性显著低于对照组（P＜0.05）。饲粮中α-亚麻酸添加水平为0.04%时,5和7日龄工蜂幼虫lysozyme和PO相对表达量显著高于对照组,但饲粮中α-亚麻酸添加水平为0.08%时,lysozyme相对表达量会显著降低（P＜0.05）。【结论】α-亚麻酸对意大利蜜蜂工蜂幼虫抗氧化活性和免疫能力有一定影响,幼虫饲粮中α-亚麻酸适宜添加水平为0.02%—0.04%。  相似文献   

Plant and spider communities benefit differently from the presence of planted hedgerows in highway verges     

Isabelle Le Viol  Romain Julliard  Louis de Redon  Nathalie Machon 《Biological conservation》2008,141(6):1581-1590

Road verges should play a crucial role as a refuge for native flora and fauna in human dominated landscapes. However, the influence of construction choices, such as plantation of woody species, on the biodiversity supported by roadsides has received little attention, although the presence of hedgerows in roadsides is likely to enhance their role as a refuge, notably for woodland species. Using standardised methods, we assessed the impact of planted hedgerows on two taxonomic groups (plants and spiders) inhabiting highway verges within an intensive agricultural landscape. We examined community richness, taxonomic and functional composition in sites with and without planted hedgerows. At the site level, the response of plant and spider communities to the presence of planted hedgerows differed markedly: hedgerows were associated with significantly higher plant richness (higher α-diversity), but similar spider richness. Plant communities in sites without hedgerows appeared as a subset of communities in sites with hedgerows, whereas spider communities in non-planted sites were complementary to that of planted sites (increased β-diversity). The presence of planted hedgerows was also associated with increased taxonomic and functional trait diversity at the landscape level (γ-diversity), through an increased β-diversity in both plants and spiders. Our results thus suggest that a mosaic of planted hedgerows and grassland habitats is crucial for the maintenance of biodiversity at a landscape scale. By providing information for road practitioners and policy makers regarding their potential impact on biodiversity, these results have important direct implications for the management of road networks.  相似文献