共查询到18条相似文献，搜索用时 218 毫秒
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旨在探明豪猪血蜱卵原生质蛋白成分,为探索干扰蜱类胚胎发育和新型控蜱策略奠定基础。采用FASP法消化豪猪血蜱卵蛋白提取物,LC/MS/MS法对其进行分析、检测,搜索褐黄血蜱卵巢转录组文库、唾液腺转录组文库和中肠转录组文库,分别检出特异性肽段359、312、357条,鉴定多肽108、170、161条,终而鉴定高可信蛋白103种。豪猪血蜱蜱卵原生质中富含酶类、蛋白酶抑制剂、转运蛋白、细胞骨架蛋白、蛋白质合成与修饰、分泌蛋白以及未鉴定的蛋白共8类蛋白。 相似文献
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本研究旨在探明4个时间段褐黄血蜱（采自刺猬体表）中肠蛋白质成分,揭示参与血餐消化的蛋白种类及其含量变化规律。采用液相色谱-串联质谱法（LC-MS/MS）对采集于刺猬体表的4个时间段褐黄血蜱的中肠蛋白组成分进行检测。基于该蜱的唾液、中肠转录组翻译文库及Uniprot数据库,利用软件Mascot 2.2对所获肽段及蛋白质进行鉴定。结果显示：在褐黄血蜱中肠蛋白提取液中共检测出特异性肽段3 046条,鉴定303种蛋白,其中271种为高可信蛋白;在所有的高可信蛋白中,23种含量较为丰富,125种在后期消化阶段（第2至第4阶段）的含量为0,123种的含量发生不同程度的变化（24种蛋白的含量变化明显,其中12种含量上升明显,12种含量下降明显）。确定148种高可信蛋白来自于刺猬血清,推断24种可信蛋白可能参与蜱虫对血餐的消化。 相似文献
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在实验室条件下,对嗜群血蜱的生活史进行了观察.研究分为2组,试验组宿主为家兔,嗜群血蜱在兔体完成一个世代需要(120.6±14.5)d;幼蜱的吸血前期、吸血期、蜕皮前期和蜕皮期分别为(7.1±0.9)、(5.6±1.1)、(11.8±2.0)和(12.5±1.6)d;若蜱的吸血前期、吸血期、蜕皮前期和蜕皮期分别为(5.9±1.1)、(3.9±0.6)、(9.8±2.3)和(12.3±0.4)d;成蜱吸血前期、吸血期、产卵前期、产卵期和死亡期分别为(7.6±1.2)、(4.9±0.6)、(9.2±1.7)、(16.0±3.2)和(5.8±0.5)d;卵的孵化期为(24.2±0.9)d.而对照组以家兔、狗和牛分别作为嗜群血蜱幼蜱、若蜱和成蜱的宿主,完成其生活史需要(130.2±12.8)d,幼蜱的吸血前期、吸血期、蜕皮前期和蜕皮期分别为(7.1±1.2)、(5.8±0.8)、(12.1±2.5)和(12.3±1.7)d;若蜱的吸血前期、吸血期、蜕皮前期和蜕皮期分别为(6.2±0.7)、(4.6±0.3)、(12.4±2.1)和(12.9±0.3)d;成蜱吸血前期、吸血期、产卵前期、产卵期和死亡期分别为(7.6±1.1)、(6.1±0.4)、(10.7±0.8)、(15.7±2.4)和(8.1±0.7)d;卵的孵化期为(23.3±1.6)d.试验组与对照组相比,嗜群血蜱幼蜱和若蜱饱血体质量差异显著(P<0.05),而成蜱的饱血体质量差异极显著(P<0.01).雌蜱产卵量与饱血体质量之间呈显著正相关r=0.912(P<0.01),试验组生殖效率指数REI为10.48,生殖适合度指数RFI为6.82,均与对照组差异显著(P<0.05).本研究的结果表明兔是嗜群血蜱较适宜的宿主. 相似文献
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《畜牧兽医科技信息》2015,(7)
<正>蜱是一类以吸血为生的体外寄生虫,对人和动物具有直接和间接双重危害,但迄今为止人们尚无安全、高效、环保的防蜱灭蜱措施。以Bm86抗原为基础的抗蜱疫苗的成功研制和商品化激发了蜱学工作者鉴定功能蛋白、筛选候选抗原的热情,且成效明显。蜱唾液中含有大量的功能蛋白,对蜱吸血活动至关重要,其中一些蛋白具有疫苗开发价值,是分离、鉴定蜱唾液功能蛋白的重要来源。由于蜱个体较小,采集比较困难,制约了很多科研人员的研究进程。近两年,笔者在实践 相似文献
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亚洲璃眼蜱唾液腺差异表达基因文库的构建及分析 总被引：4，自引：0，他引：4
从单雌蜱克隆群中挑选未吸血雌蜱60只，随机分成两组。未吸血组直接剖取唾液腺．半饱血组于蜱吸血第5d采集，分离唾液腺。Trizol法提取总RNA，经第一链合成、LD—PCR、RasⅠ酶切、接头连接和抑制消减杂交（SSH）等步骤，获得差异表达基因的cDNA片段。将纯化的cDNA片段与pGEMT—Easv我体连接，转化DH5a，获得204个白色菌落。扩增检查表明，136个克隆含有插入片段，片段大小为250bp-850bm，测出有效序列120个。由10个cDNA片段的RT—PCR检查结果初步断定，本研究消减效果良好。由网上资源分析得：21个片段与其他蜱的基因，19个片段与按蚊、库蚊、钩虫、奥斯特线虫等其他吸血寄生虫的基因，9个与果蝇的基因具有同源性。 相似文献
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将吸血诱导的亚洲璃眼蜱唾液腺差异表达基因GP29的开放性阅读框插入pGEX-4T-1，转化BL21，表达出一相对质量为53ku的蛋白，其大小与预计结果一致。蛋白质的肽质量图谱和“1381．7511”肽段序列两方面的结果证明，SDS-PAGE胶板上相对质量为53ku的蛋白就是由目的基因表达的GST融合蛋白。该蛋白与Swiss PROT库中的任何蛋白均有较大差别，可能是一个新功能分子。 相似文献
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为了进行抗蜱和蜱传病疫苗的研究,本实验对亚洲璃眼蜱雌蜱吸血前后唾液腺消减文库中获取的一个全长编码基因P18进行了研究。该基因全长519bp,共编码170个氨基酸,分子量为18.36Ku,等电点为4.28。BLAST分析表明,该基因预测的氨基酸与肩突硬蜱、篦子硬蜱唾液腺的抗凝血小肽有30%～40%低度同源性。将该基因亚克隆到pET-32a( )表达载体,转化BL21(DE3)宿主菌,经IPTG诱导,重组融合蛋白以可溶性形式高效表达。将可溶性重组蛋白免疫小鼠后获得的抗血清。经免疫印迹分析表明,该重组蛋白抗体可特异性的识别半饱雌蜱唾液腺中的天然蛋白抗原,而未吸血雌蜱唾液腺中则不显现特异条带。RT-PCR结果进一步证实,该基因在蜱吸血后的唾液腺中差异表达。 相似文献
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在不同饲喂方式和温度条件下,观察我国北方长角血蜱(Haemaphysalis longicornis)甘肃株的生活史及对其生物学特性的影响。将试验蜱分组后置于不同饲喂方式和温度下,待其发育到下一阶段,叮咬实验动物,连续调查其发育阶段的生物学特性。研究发现,完成一个世代需101 d～148 d。成年饥饿雌蜱吸血期具有两个显著特征:缓慢吸血和快速吸血;经兔体叮咬的成年雌蜱其饱血体重小于羊体。不同饲喂条件下长角血蜱饱血体重、产卵量差异极显著(P0.01);在一定温度范围内,幼蜱、若蜱的蜕变时间随温度的升高而缩短;当饱血雌蜱处于发育阶段时,产卵前期、产卵期及卵的孵化期均随着温度的升高而缩短;16℃～30℃条件下长角血蜱产卵量没有变化,利于蜱的存活;4℃以下和45℃以上饱血蜱及其后代不能存活;16℃以下蜱的发育周期延长。结果表明,长角血蜱为三宿主蜱;雄蜱可以帮助雌蜱吸血;在羊体上的吸血能力优于在兔体上的吸血能力;温度对长角血蜱的发育繁殖有影响。 相似文献
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灌注寡肽与游离氨基酸对来航公鸡氨基酸吸收及循环中肽的影响 总被引：20，自引：0，他引：20
试验比较研究了在麻醉条件下来航公鸡十二指肠灌注酪蛋白水解物寡肽（COP）与游离氨基酸（FAA）对门静脉血液中氨基酸和肽的影响。结果表明：灌注后10分钟，COP组门静脉总氨基酸（TAA）含量显著地（P＜0．05）高于相同组成的等摩尔浓度的FAA1组和等百分浓度的FAA2组。FAA2与FAA1组相比，一些氨基酸含量有升高的趋势，但仅个别氨基酸显著（P＜0．05）高于FAA1组。COP组门静脉血浆游离氨基酸含量，除赖氨酸、色氨酸、酪氨酸、脯氨酸、组氨酸外，其它氨基酸都显著（P＜0．05）或极显著（P＜0．01）高于FAA1和FAA2组。在饥饿状态及灌注COP和FAA1、FAA2后，门静脉血浆中肽结合氨基酸（PBAA）分别占39．23％，35．63％，45．89％和52．63％。其中谷氨酸、丝氨酸、丙氨酸、甘氨酸比例较高，它们与其它一些氨基酸存在显著（P＜0．05）或极显著（P＜0．01）正相关。GPLC分析结果显示：灌注COP和FAA后门静脉血浆的肽量均极显著地（P＜0．01）高于饥饿对照状况，COP组鸡门脉血浆中总量和一些肽量显著高于FAA组（P＜0．01）。试验证实，鸡村寡肽的氨基酸吸收快于游离氨基酸，一部分可以完整的形式吸收进入血液循环。给予肠道FAA也会改变循环中的肽量。 相似文献
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Immunological characterization of breakdown peptides of the 104 kilodalton hemolysin of Actinobacillus pleuropneumoniae serotype 1 总被引：1，自引：0，他引：1
The 104 kilodalton (kDa) hemolysin of Actinobacillus pleuropneumoniae serotype 1, strain CM-5 was precipitated from RPMI-1640 culture supernatant using ammonium sulfate to 80% saturation. In immunoblots, a rabbit polyclonal antiserum against the 104 kDa hemolysin protein, recognized not only the original 104 kDa monomeric form of the hemolysin but other proteins in the crude antigen mixture ranging in molecular mass from 43 to greater than 125 kDa. The antiserum was able to crosslink these proteins to active hemolysin in RPMI-1640 culture supernatant resulting in bands of hemolysis in blood agar used in a contact assay. Corresponding to these bands of hemolysis, denatured peptides with molecular masses of 51, 85, 104 and greater than 125 kDa were excised and injected into rabbits. In immunoblots, the resultant antibodies recognized the injected peptide and the monomeric 104 kDa protein. However, only the rabbit antisera produced against the 104 and 125 kDa proteins contained antibodies which neutralized the active 104 kDa hemolysin in culture supernatant. These results indicate that (i) the 104 kDa protein hemolysin can exist in a higher molecular weight aggregate (greater than 125 kDa) but can also break down to peptides which have molecular masses smaller than the 104 kDa parent molecule and (ii) while several epitopes are present in the hemolysin molecule, there seems to be a restricted number of antigenic determinants responsible for inducing neutralizing antibodies and these seem to reside only in the 104 kDa parent molecule. This may have consequences, in terms of vaccine development, for the control of pleuropneumonia in swine herds. 相似文献
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Typically, in vitro methods used for estimating the amount of ileal digestible AA do not exhaustively digest samples, and arbitrary methods for separating digestible from indigestible protein are used. This may lead to over- or underestimation of digestibility coefficients. A method that exhaustively digests proteins using pepsin and pancreatin was developed, and the first objective of this research was to confirm that exhaustive digestion was indeed appropriate and to determine the fractionation method for separating digestible from indigestible proteins. For this, three homoarginine-labeled animal proteins were prepared. Samples were subsequently digested in vivo and in vitro to determine which fraction should be considered indigestible, and in vitro followed by in vivo to determine whether the extent of digestion in vivo was improved by predigestion. In vivo, soluble but unabsorbed peptides were smaller than 1 kDa, suggesting that the size of soluble peptides is not what prevents their absorption. Thus, all in vitro-soluble proteins should be considered digestible. In vitro, 88 +/- 3% of the soluble peptides were smaller than 1 kDa, with the remainder between 1 and 5 kDa, suggesting that in vitro digestion is less complete. Predigested samples were digested in vivo to the same size distribution as the nonpredigested samples. The second objective was to test whether in vitro digestibility assays based on these principles equaled in vivo digestibility. For this, digestibility data for 25 animal proteins were compared. Results showed a lack of correlation between lysine digestibility coefficients; however, across samples, the extent of digestion did not differ for lysine (P = 0.71), threonine (P = 0.26), methionine (P = 0.18), or valine (P = 0.66), whereas in vitro digestibility coefficients were lower for (the less water-soluble) histidine (P = 0.05), isoleucine (P < 0.01), leucine (P < 0.01), and phenylalanine (P = 0.05). In conclusion, in vitro digestibility assays should exhaustively digest proteins to mimic in vivo digestibility. All in vitro-soluble peptides could be considered digestible, because in vivo, no large soluble peptides were observed whose size prevented them from being absorbed. However, an in vitro assay based on these principles lacked precision for highly water-soluble AA, and underestimated digestibility for other AA. Better solubilization of the digesta and more replicates may improve the in vitro assay further. 相似文献
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【目的】 探明褐黄血蜱卵蛋白成分及其在胚胎发育过程中的作用，为筛选抗蜱生殖疫苗分子奠定基础。【方法】 提取褐黄血蜱孵化0、7、14、21 d的卵蛋白，以过滤器辅助样品制备（filter-aided sample preparation，FASP）法酶解卵蛋白，液相色谱-串联质谱法（liquid chromatography-tandem mass spectrometry，LC-MS/MS）分离、检测酶解特异性肽段，搜索该蜱中肠转录组文库鉴定卵原生质中的蛋白，登录NCBI对各蛋白进行注释；以Maxquant计算各蛋白的iBAQ （intensity-based absolute quantification）和非标记定量（Label-free quantification，LFQ），分析各蛋白在不同孵化时间的相对丰度及其增减，并对高可信蛋白序列进行GO功能富集分析。【结果】 本研究共检出特异性肽段1 044条，由此鉴定多肽291种，其中高可信的（unique peptides≥2）112种，但仅39种有相关文献。基于文献，按照功能将39种蛋白归类于酶、酶抑制剂、转运蛋白、热休克蛋白、细胞骨架蛋白及未知蛋白6类；酶、酶抑制剂、转运蛋白、热休克蛋白都有家族蛋白共存的情况。定量分析显示，在新产褐黄血蜱卵中，丰度最高的是Contig14782，其次为Contig6575。至孵化结束时，在20种高丰度蛋白中，1种（Contig2242）显著升高（P<0.05），14种显著下降（P<0.05），5种增减不显著。GO功能富集分析表明，本次鉴定的高可信蛋白主要定位于胞外区域，参与免疫调节过程，发挥结合蛋白质和RNA的作用。【结论】 褐黄血蜱卵蛋白成分复杂、种类众多，包括酶、酶抑制剂、转运蛋白、热休克蛋白、细胞骨架蛋白等，其中酶是最多的一类。在孵化期间，绝大多数蛋白酶和酶抑制剂丰度显著下降。 相似文献
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B C Simpson M S Lindsay J R Morris F S Muirhead A Pollock S G Prichard H G Stanley G R Thirlwell A G Hunter J Bradley 《The Veterinary record》1987,120(6):135-138
Five groups of Tswana-cross castrated male cattle between 20 and 30 months of age (a total of 158 animals) were transported from a ranch in a heartwater-free area of south Botswana to a feedlot near Gaborone in the east of Botswana where heartwater is endemic. On arrival, one group was vaccinated intravenously with the Onderstepoort sheep blood heartwater vaccine, one group was vaccinated intravenously with the new Onderstepoort tick-derived heartwater vaccine and a third group was vaccinated subcutaneously with this tick-derived vaccine. Vaccine reactions were blocked with long acting oxytetracycline on the first day of fever. A fourth group had a series of injections of long acting oxytetracycline on days 0, 7, 14 and 21 after arrival, and a fifth served as untreated controls. The animals remained at the feedlot for 65 days during which time they faced a low level of challenge by Amblyomma hebraeum ticks. None contracted heartwater and so they were then challenged, together with a further group of control cattle, with a dose of the sheep blood vaccine. Some animals in all groups had severe heartwater reactions and died despite therapy, but 76.7 per cent, 64.5 per cent and 74.3 per cent of the cattle in the blood vaccine, intravenous tick vaccine and long acting oxytetracycline groups respectively were resistant to challenge, compared with 48.3 per cent of the subcutaneous tick vaccine group and 36.4 per cent of the controls. It was concluded that intravenous vaccination of susceptible adult cattle with either the blood or the tick-derived vaccine needs careful monitoring in the month after vaccination and does not necessarily result in immune animals.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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Patgaonkar M Aranha C Bhonde G Reddy KV 《Veterinary immunology and immunopathology》2011,139(2-4):176-186
Antimicrobial peptides (AMPs) serve as a first line of host defense and represent an important, though poorly understood components of the innate immune system. The present study was an attempt to identify and characterize the major molecules having anti-bacterial activities from the vaginal fluid of rabbit, Oryctologus cuniculus. AMPs from the rabbit vaginal fluid (RVF) were identified in the acid extracts of pooled RVF samples after RP-HPLC purification. The protein, RVFAMP was effective against gram negative (Escherichia coli and Pseudomonas aeruginosa) and gram positive (Staphylococcus aureus and Streptococcus pyogenes) bacteria. The results of acid urea-PAGE-gel overlay assay (AU-PAGE-GOA) demonstrated clear zone of growth inhibition of E. coli corresponding to 6 and 15 kDa protein bands. LC-MS data of these proteins indicated that 15 kDa protein consists of lysozyme, lipopolysaccharide binding protein (LBP), hemoglobin-α and β subunits (Hb-α/β), whereas 9 kDa protein band consists of transthyretin and calcyclin while uteroglobulin and neutrophil antibacterial peptide-5 (NAMP-5) are present in the 6 kDa protein band. Of the eight proteins, Hb-α derived protein was further characterized, as it showed the highest Probability Based Mowse Score (PBMS) of 288. A 25mer peptide, RVFHbαp was active against several clinical pathogens as demonstrated by minimum inhibitory concentration (MIC) and radial diffusion assays (RDA). The interaction of RVFHbαP with bacterial liposome membrane was assessed by calcein dye leakage assay. RVFHbαP did not show cytotoxicity against human endocervical cells (End1/E6E7) or erythrocytes. RT-PCR and immunofluorescence results revealed the expression of RVFHbαP mRNA and protein in rabbit vaginal tissue. To the best our knowledge, this is the first report describing the detection of AMPs in RVFs. In conclusion, these studies indicated that vaginal epithelial cells (VEC) derived RVFHbαP may have therapeutic potential in the management of reproductive well being of rabbits. The major reason for undertaking this study in rabbits is that, it forms an excellent in vivo model system for human's studies. 相似文献
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本研究利用生物信息学分析日本血吸虫编码Akt蛋白的cDNA并对编码该蛋白的cDNA进行了克隆和原核表达。生物信息学分析表明日本血吸虫存在两个Akt蛋白。利用PCR将其中一个Akt蛋白催化功能域的编码cDNA进行了克隆,并利用原核表达系统对此蛋白片段进行诱导表达并进行了重组蛋白的纯化,将重组蛋白免疫新西兰大白兔制备了多克隆抗体。Western blot分析表明,该抗体能被日本血吸虫Akt重组蛋白特异性识别。本研究为进一步研究该蛋白的功能奠定了基础。 相似文献