首页 | 官方网站   微博 | 高级检索  
相似文献
 共查询到19条相似文献,搜索用时 218 毫秒
1.
γ-射线及大肠杆菌诱导蓖麻蚕产生抗菌物质的研究   总被引:3,自引:1,他引:2  
戴祝英  吴冬秀 《昆虫学报》1989,32(3):271-277
1.用γ-射线辐照和用大肠杆菌处理蓖麻蚕五龄幼虫和蚕蛹均能诱导血淋巴产生抗菌物质,两种诱导源诱导所得活性物质的抗菌活性相似.2.研究了五龄幼虫期和蛹期诱导产生抗菌物质的动力学,发现五龄幼虫在饷食后第2—4天诱导活力最高,产生抗菌物质的持续时间亦较长;蛹期则从化蛹当天至第4天之间诱导活力较高,并可持续15天左右,高峰期一般在诱导后2天至4天之间.3.对诱导后的蓖麻蚕血淋巴进行了电泳测活和葡聚糖凝胶初步分离,发现不论幼虫期或蛹期至少可得三个活性组分,其中既有类似于P5的大分子抗菌物质,也有类似于P9A和P9B的抗菌多肽;并首次发现一种分子量约70000—75000道尔顿的新抗菌蛋白.  相似文献   

2.
蜚蠊灭菌肽的诱导及初步分离分析   总被引:13,自引:1,他引:12  
张然  陈濂生 《昆虫学报》1990,33(1):7-13
昆虫经诱导盾产生灭菌肽的研究近年来已有很大进展,有关这方面的研究工作绝大多数都是以有翅亚纲内生翅类鳞翅目(主要是蚕类)昆虫和少数双翅目昆虫为材料.本文首次以有翅亚纲外生翅类蜚蠊目的美洲蜚蠊(Ptriplaneta americana L.)为实验昆虫,用Escherichia coil K12 strain D31作诱导源,对不同发育期、不同性别、不同成虫期的蜚蠊进行诱导后,采用含菌培养基平板测活方法,就存在个体数及能产生抗菌物质的个体数进行了初步的研究.发现成虫日龄在10天之内的雄性蜚蠊能够产生抗菌物质的个体百分比最高.抗菌物质出现的高峰期是在诱导后第三、四天.用滴滴涕和溴氰菊酯作诱导源对雄性蜚蠊的诱导实验表明,杀虫剂也能诱导蜚蠊产生抗菌物质,而且所诱导产生抗菌物质的活性强度(用抑菌圈直径表示)高于大肠杆菌所诱导的.滴滴涕和溴氰菊酯的重复诱导可提高蜚蠊产生抗菌物质个体百分比.蜚蠊经诱导后产生的抗菌物质具有广谱性,对苏云金杆菌、金黄色葡萄球菌、枯草杆菌及绿脓杆菌等有较强的抗菌活性,而对大肠杆菌D31、大肠杆菌、粘质沙雷氏杆菌和溶壁微球菌等有较弱的抗菌活性.用肽类物质的指纹图谱法分离蜚蠊血淋巴抗菌物质,发现经诱导后血淋巴中确有新的肽类物质产生,该物质具抗菌活性,用DABITC法分析,其N-末端氨基酸为赖氨酸.  相似文献   

3.
大肠杆菌D31诱导柞蚕蛹产生抗菌多肽。同时,溶菌酶和凝集素活性都比诱导前有明显增高.其活力高峰、抗菌多肽在第7天左右、溶菌酶在第5天,而凝集素在第3天即达最高水平。不同品种的柞蚕蛹,经诱导产生的三种活性物质其活力差异不明显,但741、河四和小混品种中的抗菌多肽P9A及P9B组成比例较高。上述三种活性物质的诱导变化与性别有关,雄性高于雌性。比较了柞蚕蛹和家蚕经细菌诱导后上述三种活性物质的变化,家蚕凝集素活力很低,诱导后活力增高不明显。抗菌活力及溶菌酶活力的提高程度柞蚕也高于家蚕。聚肌胞核苷酸(Poly I:C)也能诱导两种蚕产生抗菌多肽及溶菌酶,但活力提高的显著程度都不及大肠杆菌诱导,凝集素活力变化也不显著。  相似文献   

4.
【目的】Toll信号通路是昆虫中重要的免疫信号通路,其中Toll受体在保持Toll通路的正常免疫应答、抵抗外源病原体中起到关键的作用。本研究旨在探究肽聚糖和金黄色葡萄球菌Staphylococcus aureus对家蚕Bombyx mori Toll受体基因BmToll9-1和BmToll9-2表达的影响。【方法】将革兰氏阳性菌细胞壁主要成分肽聚糖和革兰氏阳性菌金黄色葡萄球菌分别注射感染家蚕5龄第1天幼虫,诱导其发生免疫反应;采用实时荧光定量PCR分析注射后不同感染时间点BmToll9-2和BmToll9-1基因在家蚕幼虫中肠、表皮、脂肪体和丝腺中的相对表达水平。【结果】往家蚕5龄幼虫中注射肽聚糖或金黄色葡萄球菌后,BmToll9-2基因出现了时间和组织的差异性表达。注射肽聚糖和金黄色葡萄球菌均能诱导5龄幼虫中肠BmToll9-2基因的表达上调,注射肽聚糖和金黄色葡萄球菌分别在3和6 h时对基因表达的诱导效果最好,且注射金黄色葡萄球菌比注射肽聚糖对基因表达的诱导效果更好。注射金黄色葡萄球菌能引起5龄幼虫表皮、脂肪体和丝腺中BmToll9-2基因的表达上调,分别于注射后24, 6和24 h时诱导效果最好。注射金黄色葡萄球菌亦能诱导同源的BmToll9-1基因的上调表达。【结论】家蚕幼虫BmToll9基因在肽聚糖或金黄色葡萄球菌注射处理后均能在不同组织中发生上调表达,推测BmToll9基因参与了家蚕对肽聚糖和金黄色葡萄球菌的免疫反应。  相似文献   

5.
华北大黑金龟幼虫对注射细菌的免疫反应   总被引:2,自引:0,他引:2       下载免费PDF全文
杨明华 《昆虫学报》1985,(2):160-164
在华北大黑金龟三龄幼虫体腔内分别注射病原菌Bacillus popilliae及非病原菌Bacillus megaterium,观察血细胞总数(THC)和血淋巴蛋白质组分的变化。注射后15分钟,两者使THC都下降50%左右,1小时后开始回升。这时,注射非病原菌的幼虫THC上升到最高峰,约超过正常的86%。此后10小时内又经过三次下降和回升。用聚丙烯酰胺凝胶电泳测定表明,注射病原菌的幼虫血淋巴会产生蛋  相似文献   

6.
油松毛虫幼虫抗菌物质及其抗菌活性研究   总被引:1,自引:0,他引:1  
采用体内注射法,对油松毛虫Dendrolimus tabulaeformis Tsai et Liu 3龄幼虫注射浓度为1.0×10^8孢子/mL的大肠杆菌Escherichia coli菌悬液,诱导其产生抗菌物质,12h后收集制备血淋巴粗提液。分别测定对4种细菌和4种真菌的抗菌活性,并检测温度、pH值变化和反复冻融对抗菌活性的影响。采用Tricine—SDS—PAGE电泳法确定抗菌物质的分子量大小。结果发现,诱导后的油松毛虫3龄幼虫血淋巴粗提液对试验的细菌和真菌均有不同程度的抑制作用,其中对革兰氏阴性细菌大肠杆菌E.coli和变形杆菌Proteus species的抑制作用强于革兰氏阳性细菌金黄色葡萄球菌Staphylococcus aureus和枯草芽孢杆菌Bacillus subtilis。血淋巴粗提液的的抗菌活性在50℃水浴处理和4~9的pH值条件下保持稳定,反复冻融1~5次抗菌物质的活性降低4.3%~14.9%。电泳结果显示,诱导组血淋巴粗提液在23.2kD和15.0kD处分别出现一条特异性条带,在60.4kD处蛋自带变浅。由此推测从油松毛虫幼虫体内诱导产生的抗菌物质分子量可能是23.2kD和15.0kD。  相似文献   

7.
中蜂抗菌物质的诱导   总被引:2,自引:0,他引:2  
陈学新  陈卫良 《昆虫知识》1999,36(4):215-218
本文报道了用大肠杆菌注射处理中蜂Apis cerana Fabricius后,用含菌培养基平板测活法,就中蜂抗菌物质的产生规律及抗菌活性进行了初步研究。结果表明,不同诱导源均可诱导中蜂产生抗菌物质,但诱导的抗菌物质的抗菌活性则有一定的差异。用大肠杆菌重复诱导则使中蜂的成活率和抗菌物质的抗菌活性有很大提高。注射大肠杆菌后冲蜂抗菌物持产生高峰在48小时左右。诱导后产生的抗菌物质具有广谱性,对大肠杆菌Ascherichia coli、枯草芽孢杆菌 Bacillus  subtilis、苏云金杆菌 Bacillus thuring-iensis、巨大芽孢杆菌 Bacillus megatherium、黑腐菌 Xanthomonas campestris 等均有抑菌作用,而对真菌白僵菌Beauveria bassiana未发现抑菌作用。中蜂麻醉方法以冷冻法最为简便、易行。  相似文献   

8.
黄粉甲幼虫抗菌物质的诱导及其抗菌活性   总被引:19,自引:2,他引:17  
采用饥饿法、紫外线照射法和针刺法处理黄粉甲Tenebriomolitor 6龄幼虫后均能诱导其 产生抗菌物质,收集的血淋巴上清液对真菌有抑制作用,对细菌无抑制作用;经热处理后的血 淋巴上清液则对细菌有抑制作用,而对真菌无抑制作用。SDS-PAGE检测结果发现,与未诱导的 对照相比经诱导的黄粉甲幼虫血淋巴中,原有的一类大分子蛋白质如分子量分别为97kD、44 kD和37 kD左右的蛋白质缺失;而ESI-MS分析结果显示诱导后比诱导前黄粉甲幼虫血淋巴中有 小分子物质产生,推测可能是此类缺失蛋白质分解为小分子量的抗菌肽,从而表现出抗菌活性 。  相似文献   

9.
大头金蝇幼虫免疫系统对大肠杆菌诱导的应答研究   总被引:11,自引:2,他引:9  
用大肠杆菌作为诱导菌,使得大头金蝇幼虫产生抗菌物质及凝集素,其活力高峰分别在第48和36小时。诱导后第48小时血淋巴稀释100倍,对金黄色葡萄球菌的致死率仍达18%。诱导后36小时血淋巴中凝集素滴度达512。聚丙烯酰胺电泳显示诱导后的血淋巴中新增4条新带,并有2条染色明显加深的条带。  相似文献   

10.
观察不同生长期家蚕幼虫血淋巴在体外的黑化速度和对大肠杆菌生长的影响结果显示,随食桑生长幼虫血淋巴的黑化速度逐渐变快,对大肠杆菌生长的抑制作用逐渐增强。RT-PCR实验显示,黑色素合成催化酶Bm Tan、Bm Po-1、Bm Yellow-f和Bm Ddc等的基因在家蚕5 L 3 d血淋巴中表达量高,Bm Black、Bm Yellow和Bm Pah等的基因也有明显表达。q PCR分析显示,黑化病蚕中Bmtan、Bmddc、Bmyellow、Bmebony和Bmblack,尤其Bmddc表达发生了显著上调。与对照相比,Ddc酶的抑制剂能显著抑制脂多糖对血淋巴的诱导黑化作用。用大肠杆菌注射家蚕幼虫,血淋巴中多巴和多巴胺的含量明显上升。这些表明家蚕幼虫血淋巴黑化与防御免疫有关,Bmddc很可能在幼虫血淋巴的免疫黑化中发挥作用。  相似文献   

11.
柞蚕滞育蛹的体液防卫反应   总被引:5,自引:0,他引:5  
任淑仙  远立红 《昆虫学报》1996,39(4):354-359
本文对柞蚕Antheraea pernyi滞育蛹体液防卫的某些特性做了报道。结果表明:(1)经注射大肠杆菌诱导6h后,体液中出现了抗菌活性物质。诱导7d后,抗菌活力达到峰值,15d后活力消失。用生理盐水诱导的反应较快,4d后达到峰值,10d后活力消失。(2)经大肠杆菌诱导后的体液对苏芸金杆菌、大肠杆菌、枯草杆菌及金黄葡萄球菌均有杀菌效应。在最初5min内杀菌效率较高。注射生理盐水诱导的体液较大肠杆菌诱导的杀菌效果低。(3)用昆虫病原菌苏芸金杆菌及非病原菌大肠杆菌作不同的诱导源,诱导1d后的体液对大肠杆菌的抗菌活力差别不显著,但对柞蚕蛹的生活力及发育有明显影响;用苏芸金杆菌诱导的柞蚕滞育蛹3d后死亡,用大肠杆菌诱导的蛹能正常羽化。(4)在一定范围内用不同剂量的大肠杆菌进行诱导,其抗菌活力不受影响,但超过一定限度,既使是非病原菌也可以突破昆虫的防卫功能。  相似文献   

12.
家蚕抗菌肽CMIV基因结构改造及表达产物的研究   总被引:20,自引:0,他引:20  
参照天然抗菌肽CMIV组分的氨基酸序列,作了近50%的改动,根据大肠杆菌偏爱的密码子,设计并人工合成了抗菌肽基因片段.将人工合成的抗菌肽类CMIV基因先重组到测序载体pUC118上,经过序列分析,发现克隆于载体pUC118上的基因片段与设计的序列完全一致.再将该基因片段重组到表达载体pET28(a)上,抗菌肽以融合蛋白的形式表达.融合蛋白经镍-金属离子胶亲和层析纯化后,再用CNBr裂解,最终产物具有与天然抗菌肽相同的生物学活性  相似文献   

13.
The immune system in the Chinese oak silk moth, Antheraea pernyi, has been compared with that of the Cecropia moth which has been characterized earlier. Antibacterial activity against Escherichia coli was induced in diapausing pupae by injection of viable E. coli or Enterobacter cloacae. The activity reached a maximum on day 7-8 after which the response gradually declined. The pupae produced a set of immune proteins with P4 and P5 as major labelled components similar to that earlier found in Cecropia. The major antibacterial factor in A. pernyi was cecropin D. A procedure is described for the isolation of cecropin B and D, which is in principle similar to the one used for the isolation of the corresponding cecropins from Cecropia pupae. Amino acid sequence analyses of the A. pernyi cecropins show the D form to contain 36 amino acid residues and that both cecropins have blocked C-termini. The general structure of cecropins having a charged N-terminal region (residues 1-21) followed by a long hydrophobic stretch (residues 22-32) is well conserved. Cecropin B and D from A. pernyi differ from the corresponding proteins in Cecropia by four and three conservative amino acid replacements, respectively. The homology between the cecropins from the two insects suggests that they orginate from a single ancestral gene. The antibacterial activity was tested against nine different bacterial species. Evolutionary aspects of the cecropins are discussed.  相似文献   

14.
The present study elaborates a cost-effective and transfectant-free method for generating recombinant Bombyx mori (silkworm) nucleopolyhedrovirus in silkworm larvae and pupae by injecting invasive Escherichia coli carrying BmBacmid [BmNPV (B. mori nucleopolyhedrovirus)-Bacmid] into larval haemocoel. Up to 109 PFU (plaque-forming units)/ml of infective recombinant baculovirus was generated in the silkworm by intrahaemocoelic injection with 106 DAP (diaminopimelic acid) auxotrophic and BmBacmid containing E. coli cells expressing both invasin and listeriolysin. Thus 1?ml of overnight culture of E. coli is sufficient to inject more than 2000 larvae, while DAP costing up to $1 is enough to inject about 4000 larvae. Recombinant proteins can be controlled to be expressed mainly in pupae by adjusting the injection dose, too. In this new method, many original manipulations have been eliminated, including BmBacmid preparation and the subsequent complex transfection procedures. Hence it is a time- and cost-saving means for large-scale injection of B. mori for recombinant baculovirus production in comparison with the traditional transfection methods, which may play an important role in the industrial development of the BmNPV-silkworm bioreactor.  相似文献   

15.
1. Effect of oral administration of live or formalin-treated Escherichia coli (E. coli) K-12 to the fifth instar, days 1 and 3 larvae of the silkworm, Bombyx mori, on induction of antibacterial activity in the haemolymph was investigated using the silkworms reared on an artificial diet under completely aseptic conditions. 2. When live E. coli was administered to the male day 1 larvae, low but significant antibacterial activity of 3.8 mm was detectable in the haemolymph of one individual at 48 hr after immunization. The proportion of the larvae to express antibacterial activity increased thereafter and at 120 hr after immunization, all three individuals showed antibacterial activity. In day 3 male larvae, activity was detectable at 48 and 72 hr after immunization. 3. When formalin-treated E. coli was orally administered to days 1 and 3 male larvae, no activity was detectable at any time post-immunization. 4. In the second experiment, when day 1 larvae, females and males were orally immunized with live E. coli, only females showed antibacterial activity in the haemolymph, beginning from 24 hr after immunization and up to 96 hr. 5. Removal of an antibiotic, chloramphenicol, from ingredients of an artificial diet was required for induction of antibacterial activity with oral administration of live E. coli. 6. When live E. coli that grows at pH 9.0 was selected and used for oral immunization, antibacterial activity was induced both in females and males at 72 hr after immunization and the activity was observed at 96 hr. 7. These results suggest that establishment of oral immunization with live E. coli in the silkworm larvae requires multiplication of E. coli in the midgut lumen and possibly its colonization on the luminal surface.  相似文献   

16.
比较了不同诱导源对诱导柞蚕蛹血淋巴中抗菌活力的动力学变化。各种化学试剂包括生理盐水都能诱导产生抗菌物质。在不引起死亡的刺激量下,刺激量愈大,诱导的抗菌活力愈高。不同金属络合剂所诱导产生的抗菌物质组份可有很大区别。如二氮杂菲使P_(9E)及P_(9A)的组份比例增高,而EDTA则可提高P_(9B)及P_(9D)组份的比例。P_(9E)和P_(9A),P_(9D)和P_(9B)的产生分别有对应关系,讨论了是否由两个不同基因所控制。 大肠杆菌诱导后的柞蚕蛹,除在血淋巴中测得抗菌活力,在雌雄的生殖腺内含物中也能测得。摘除生殖腺后虽然在血淋巴中仍能诱导出抗菌物质,但应答较慢,活力也较小,也可能由于手术创伤较大,同样的刺激量容易引起死亡。  相似文献   

17.
Immune hemolymph was collected from fifth instar larvae and 1-day-old pupae of Heliothis virescens after injection of prepupae with live Enterobacter cloacae. Induction of antibacterial activity against Escherichia coli K12 D31 was 7.5 times greater in pupal than in larval immune hemolymph. Lysozyme activity of immune pupal hemolymph against Micrococcus lysodeikticus was 11 times greater when compared with lysozyme activity of immune larval hemolymph. Early pupal immune response with regard to antibacterial activity was much greater than larval immune response in H. virescens. Normal pupal hemolymph showed an increase in antibacterial activity and lysozyme that was induced during metamorphosis. Antibacterial protein was isolated together with lysozyme by gel filtration chromatography and then separated from lysozyme by sequential electrophoresis with a native acid gel and SDS gel. Molecular mass of antibacterial protein was estimated to be 12 kDa. The N-terminal amino acid sequence of 12-kDa protein was different from those of antibacterial molecules found in other insects and has not been identified before. A sample containing 12-kDa protein was negative for immunoblotting with anti-synthetic cecropin B antibody. We have named the novel 12-kDa antibacterial protein viresin. Viresin showed antibacterial activity against several Gram-negative bacteria including E. cloacae but not against Gram-positive bacteria.  相似文献   

18.
家蚕对马尾松毛虫质型多角体病毒的敏感性   总被引:2,自引:0,他引:2  
用虫体克隆技术,对马尾松毛虫质型多角体病毒湖南株(DpCPV-HN)进行了分离纯化,鉴定为质型多角体病毒1型。以家蚕春蕾×镇珠杂种F1代及自交的F2代4或5日龄幼虫进行毒力测定,以纯化的家蚕质型多角体病毒对F1代幼虫的毒力测定为对照。结果表明:家蚕品种春蕾×镇珠对家蚕质型多角体病毒敏感,马尾松毛虫质型多角体病毒湖南株能引起其感染发病;马尾松毛虫质型多角体病毒湖南株感染家蚕品种春蕾×镇珠F1代幼虫和F2代幼虫28天后的半致死剂量(LD50)分别为885个和18个CPB(质多角体),前者为后者的49倍。马尾松毛虫质型多角体病毒湖南株感染后的家蚕,其结茧率、化蛹率、羽化率、全茧量、茧层量和单蛾产卵数均有所下降,全茧量、茧层量、茧层率和单蛾产卵数与病毒感染剂量之间无显著关联。  相似文献   

19.
Many proteins, including antibacterial peptides in the hemolymph, are induced by bacterial infections. We found two bacterially inducible carboxylesterases (CEs) in the hemolymph of the silkworm, Bombyx mori. CEs Est-1 and 2 were induced by lipopolysaccharide injection after 6 hours as well as E. coli infection. We found that bacterially inducible CEs clearly differed from noninducible CEs, including juvenile hormone esterases, in pI values, migration on analytical native PAGE, and inhibitor sensitivity. We are now studying the features and functions of these CEs.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司    京ICP备09084417号-23

京公网安备 11010802026262号