家蚕IAP互作蛋白的筛选、鉴定及其对BmNPV增殖的影响

目的 细胞凋亡作为昆虫免疫应答的重要组成部分,在病毒与宿主相互作用的过程中扮演着重要角色,以细胞凋亡相关基因为研究对象对阐明其在病毒感染过程中的作用具有重要的参考价值。本研究旨在筛选家蚕凋亡抑制蛋白（Bombyx mori inhibitor of apoptosis protein,BmIAP）的相互作用蛋白,并验证其在家蚕核型多角体病毒（Bombyx mori nucleopolyhedrovirus, BmNPV）侵染过程中与Bmiap的相互调控关系及在BmNPV增殖过程中的作用,为探明宿主与BmNPV相互作用的分子机制提供理论依据。方法 利用免疫共沉淀技术筛选在BmNPV侵染家蚕细胞过程中与BmIAP相互作用的蛋白,对获得的特异性差异条带进行LC-MS/MS蛋白质谱分析,根据蛋白分子量大小并利用生物信息学方法对获得的蛋白进行鉴定,确定候选基因;通过分子克隆技术对候选基因进行克隆,利用SMART在线工具及BioEdit分别对候选基因的结构域进行预测及多序列比对分析;通过免疫荧光试验验证BmIAP和候选蛋白的细胞共定位情况,并进一步利用免疫共沉淀验证它们的相互作用;分别通过真核表达和基于CRISPR/Cas9的基因编辑系统对候选基因进行过表达和敲除,利用qRT-PCR技术检测相应基因的表达情况,以确定Bmiap和Bmpp5在BmNPV侵染过程中的调控关系;同样,在过表达和敲除Bmpp5后检测杆状病毒Vp39的表达量,以确定Bmpp5对BmNPV增殖的影响。结果 通过免疫共沉淀获得7个可能与BmIAP相互作用的宿主蛋白和1个BmNPV蛋白,进一步分析确定1个与凋亡相关的候选基因Bmpp5;Bmpp5的开放阅读框为1 473 bp,编码490个氨基酸,预测的蛋白分子量约为56 kD,含3个TRP结构域和1个PP2Ac结构域,在昆虫间具有较高的保守性;免疫荧光检测证明BmIAP和BmPP5共定位于细胞质中,免疫共沉淀结果表明两者可以相互作用;在BmNPV侵染过程中,过表达Bmiap后,Bmpp5的表达量显著上调,而敲除Bmiap后,Bmpp5的表达量显著下调,表明Bmiap能够促进Bmpp5的表达;同时,过表达Bmpp5后,Bmiap显著上调表达,而敲除Bmpp5后,Bmiap显著下调表达,表明Bmpp5同样能够促进Bmiap的表达;过表达Bmpp5能够促进BmNPV的增殖,敲除Bmpp5能够抑制BmNPV的增殖,表明Bmpp5的表达利于BmNPV的增殖。结论 鉴定了1个能够与BmIAP相互作用的蛋白BmPP5,且该蛋白在昆虫中高度保守;在BmNPV侵染过程中,Bmiap和Bmpp5能够相互促进,且Bmpp5能够促进BmNPV的复制增殖。     

Comparative analysis of survivin expression in untreated and relapsed canine lymphoma

BACKGROUND: Survivin, a member of the inhibitor of apoptosis protein family, has a dual role in tumor cell proliferative and antiapoptotic pathways. Survivin expression has been shown to be a negative prognostic factor in several cancers of humans, including B-cell non-Hodgkin's lymphoma. HYPOTHESES: High survivin expression will be a negative prognostic factor in dogs with lymphoma (LSA) treated with chemotherapy. In addition, survivin expression will be upregulated in relapsed canine LSA when compared with patient-matched, pretreatment biopsies. ANIMALS: Thirty-one client-owned dogs with stage IIIa or IVa LSA. METHODS: Retrospective evaluation of survivin immunoreactivity was performed on pretreatment lymph node biopsies and patient-matched samples obtained from dogs at relapse after being treated with an abbreviated CHOP-based protocol. RESULTS: In this population of dogs presenting with stage IIIa or IVa B-cell LSA, those dogs that had high survivin immunoreactivity scores had a significantly (P < .01, hazard ratio = 0.30) shorter median disease-free interval than did dogs with low survivin immunoreactivity scores (171 days versus 321 days, respectively). Survivin immunoreactivity was not significantly different in relapsed canine LSA when compared with patient-matched, pretreatment biopsies. CONCLUSIONS AND CLINICAL IMPORTANCE: Survivin expression is a negative prognostic factor that can predict early treatment failure of dogs that present with stage IIIa or IVa, B-cell LSA when treated with a CHOP-based protocol.     

斜纹夜蛾核型多角体病毒安徽和县株系细胞凋亡抑制相关基因的克隆与遗传变异

为分析核型多角体病毒(NPV)分离株内细胞凋亡抑制相关基因的多态性,从安徽和县采集感染病毒的斜纹夜蛾病虫,分离出斜纹夜蛾核型多角体病毒(Spodoptera litura nucleopolyhedrovirus,Splt NPV)15个活体克隆株系,研究其细胞凋亡抑制基因(IAP)和P49基因多态性。发现15个Splt NPV安徽和县克隆株系中的IAP基因序列均无差异,且与已报道的Splt NPV中国G2株的同源性达100%;Splt NPV各克隆株系中的P49基因序列也无差异,且与Splt NPV中国G2株的P49基因同源性达99%。表明Splt NPV安徽和县株系IAP和P49基因均较为保守,没有表现出遗传变异。     

Threshold change in forest understory vegetation as a result of selective fuelwood extraction in Nairobi, Kenya

We examined whether heavy fuelwood collection can cause threshold change in understory forest community and evaluated how selective wood extraction might lead to delayed forest recovery in an urban forest of Nairobi, Kenya. Piecewise regression which represents strongest support for threshold change provided the best fit for the relationships between understory floristic composition (i.e. DCA axis 1) and human disturbance gradients (i.e. canopy cover, and distance from the slum), where threshold changes were detected at c.a. 350 m from the slum and c.a. 30% canopy cover. Only one tree species significantly indicated communities beyond the threshold while an aggressive invasive alien plant (IAP) Lantana camara was strongly represented. Total species diversity along the two human disturbance gradients peaked before the threshold was reached, suggesting that decline in species diversity along the prevailing disturbance gradient might be able to forecast threshold change. Tree species richness in the understory rapidly declined as the threshold was surpassed while other growth forms (i.e. shrubs, herbs and climbers) were relatively unaffected. The effect of selective tree cutting was indirectly impacting the forest understory as species richness pattern of preferred and non-preferred species paralleled that of trees and shrubs, respectively. Thickets of L. camara can negatively affect indigenous flora and its establishment was favored under selective fuelwood extraction removing certain tree species while leaving the IAP untouched. Shading can readily eliminate the IAP, but weak tree regeneration beyond the threshold suggested forest recovery might be delayed for longer than expected because of the interaction between selective fuelwood use and the IAP.     

基于SST芯片远程升级方案的设计与实现

单片机的IAP技术可以实现应用程序控制下的在线编程,运用此技术对单片机软件在线升级,可以减少产品的维护成本,降低软件带来的风险。本文以SST公司的SST89E564RD这款单片机为例,首先介绍其IAP功能,然后在此基础之上设计了产品远程升级的硬件方案和软件方案,并针对升级过程中出现的异常情况给出了相应的处理措施。实际应用中证明通过文中的方案可以成功地对单片机产品的软件进行升级,降低了产品后期维护的困难。     

IAP对肉仔鸡生产性能的影响

将56只14日龄体重约350 g的AA雄性肉仔鸡按体重分为4组(n=14)腹腔注射胰岛激活蛋白(IAP),以研究IAP对肉仔鸡生产性能的影响。结果表明,给肉仔鸡分别腹腔注射0,0.5,1.0和2.0μg/只IAP,每只鸡4周的平均体增重分别为60.9±17.0,60.3±5.6,58.0±6.5和58.8±6.5 g/d,自由采食量分别为106.5±8.9,107.8±6.4,104.7±7.0和105.5±5.4 g/d,全净膛重分别为1360±69,1406±18,1393±37和1439±42 g/只,腹腔脂肪重分别为48±10,52±9,43±8和38±10 g/只,胸肌 腿肌重分别为620±47,645±32,657±35和688±42 g/只。各组血糖浓度分别为8.77±1.31,12.77±0.91,12.98±1.98和13.40±1.47 mmol/L,胰岛素浓度分别为10.77±1.67,13.81±3.67,15.54±3.53和13.73±5.25μU/mL,T3浓度分别为1.25±0.64,1.16±0.38,1.21±0.43和1.20±0.61 ng/mL,T4浓度分别为15.83±5.87,18.03±4.94,17.99±6.00和14.64±5.18 ng/mL。本实验表明,IAP对禽类也具有生物学作用,可同时提高肉仔鸡的血糖和胰岛素浓度,增加全净膛重和胸腿肌重,并有减少仔鸡腹腔脂肪重量的趋势,但对肉仔鸡的采食量、体增重和血液甲状腺素浓度无显著影响。     

家蚕iap基因在原核表达系统中的表达

为研究IAP蛋白抑制细胞凋亡的作用,用PCR法扩增出家蚕iap基因的cDNA,然后插入pET28a载体,得到阳性质粒pET28a-iap,并转化感受态细胞BL21,使用异丙基硫代-β-D-半乳糖苷(IPTG)诱导IAP蛋白的表达,应用SDS-PAGE和Western Blotting法分析目的蛋白,得到一条分子量约为38.8 kDa的特异表达条带,与表达产物的理论值相符。为进一步研究BmIAP蛋白的功能奠定了基础。     

基于BH20的温室无线监控系统

针对我国农业生产地域分散、偏远、现场信息获取困难以及架设有线网络成本高等特点,利用室外型无线覆盖模块(IAP)和点对点传输模块(BH20)等设备构建无线通信网络,并结合计算机控制、网络通信编程以及数据库等技术,开发了一套温室无线监控系统.测试结果表明,系统能够满足温室环境控制的要求.     

贵阳市区树木附生苔藓植物的种类及大气净度指数

为了摸清贵阳市区树木附生苔藓植物的种类,为应用苔藓植物监测贵阳市大气污染状况提供参考,采用实地考察、定点取样和标本鉴定相结合的方法,于2011年8—11月对贵阳市树木附生苔藓植物的种类、盖度、频度等指标进行调查,并分析其大气净度指数。结果表明:贵阳市区树木附生苔藓植物共19科38属79种(包括亚种、变种),苔类植物4科5属12种,藓类植物15科33属67种;大气净度指数(IAP)为0.63～18.67;大气污染情况,白云区、金阳区为清洁区,IAP最高,为10.67～18.67;云岩区、花溪区次之,IAP值为3.05～9.54;小河区、南明区、乌当区的污染相对较重,IAP值最小,为0.63～2.53;狭叶小羽藓和鳞叶藓的分布范围较广、生长较好,可作为监测贵阳市区大气污染状况的苔藓植物。