将重组质粒pET28 a-VP2转化禽多杀性巴氏杆菌弱毒株G190E40,探讨该转化菌表达的VP2是否具有免疫原性,以及重组表达质粒在转化菌内的稳定性。该转化菌在体外经IPTG或乳糖诱导,表达产物经AGP检测,VP2滴度可达1/4;经间接ELISA检测,VP2滴度...将重组质粒pET28 a-VP2转化禽多杀性巴氏杆菌弱毒株G190E40,探讨该转化菌表达的VP2是否具有免疫原性,以及重组表达质粒在转化菌内的稳定性。该转化菌在体外经IPTG或乳糖诱导,表达产物经AGP检测,VP2滴度可达1/4;经间接ELISA检测,VP2滴度可达1/512;经SDS-PAGE分析,出现VP2蛋白条带。结果表明,诱导表达的提取液中有一定量可溶性VP2。将pET28 a-VP2转化的禽多杀性巴氏杆菌弱毒株G190E40经诱导后,接种爱维茵肉仔鸡,21日龄及35日龄时,分别肌肉注射菌液0.1、0.2 m l/只(3×108个/m l活菌)。49日龄时,感染IBDV标准强毒株BC6/85。IBDVVP2血清抗体检测结果,不接种组(B)、0.1 m l接种组(C)、0.2 m l接种组(D)及空质粒转化菌接种组(E)的AGP抗体检测阳性率分别为0、65%、70%、0;间接ELISA抗体检测阳性率分别为0、80%、85%、0;IBDV强毒感染的免疫保护率分别为5%、75%、75%、5%。经t检验,B组与C组、B组与D组、E组与C组、E组与D组,P<0.05,差异均显著,但B组与E组、C组与D组,P>0.05,差异均不显著。免疫试验结果表明,诱导表达产物VP2可有效地刺激鸡体产生体液免疫应答,刺激鸡体建立的免疫力可以抵抗一定剂量强毒的攻击。在质粒稳定性测定中,第9代次及以后的3个代次,20个菌落的VP2目的基因PCR检测结果均为阴性,表明该质粒在禽多杀性巴氏杆菌弱毒株G190E40内是不稳定的。展开更多
Free fatty acids(FFAs) provide an important energy source and also act as signaling molecules.Medium to long-chain free fatty acids can activate the intracellular signal pathways in the pancreatic β-cells and play a ...Free fatty acids(FFAs) provide an important energy source and also act as signaling molecules.Medium to long-chain free fatty acids can activate the intracellular signal pathways in the pancreatic β-cells and play a role in regulating insulin secretion as an extracellular signal molecular via binding to the FFA receptor G protein-coupled receptor 40(GPR40). Furthermore,GPR40 is associated with several biological effects including cell proliferation and antiapoptosis of nerve cells.GPR40 act an important role in the connection of obesity and diabetes or cancers.GPR40 will probably become a novel kind of antidiabetic and anticancer drugs.展开更多
文摘将重组质粒pET28 a-VP2转化禽多杀性巴氏杆菌弱毒株G190E40,探讨该转化菌表达的VP2是否具有免疫原性,以及重组表达质粒在转化菌内的稳定性。该转化菌在体外经IPTG或乳糖诱导,表达产物经AGP检测,VP2滴度可达1/4;经间接ELISA检测,VP2滴度可达1/512;经SDS-PAGE分析,出现VP2蛋白条带。结果表明,诱导表达的提取液中有一定量可溶性VP2。将pET28 a-VP2转化的禽多杀性巴氏杆菌弱毒株G190E40经诱导后,接种爱维茵肉仔鸡,21日龄及35日龄时,分别肌肉注射菌液0.1、0.2 m l/只(3×108个/m l活菌)。49日龄时,感染IBDV标准强毒株BC6/85。IBDVVP2血清抗体检测结果,不接种组(B)、0.1 m l接种组(C)、0.2 m l接种组(D)及空质粒转化菌接种组(E)的AGP抗体检测阳性率分别为0、65%、70%、0;间接ELISA抗体检测阳性率分别为0、80%、85%、0;IBDV强毒感染的免疫保护率分别为5%、75%、75%、5%。经t检验,B组与C组、B组与D组、E组与C组、E组与D组,P<0.05,差异均显著,但B组与E组、C组与D组,P>0.05,差异均不显著。免疫试验结果表明,诱导表达产物VP2可有效地刺激鸡体产生体液免疫应答,刺激鸡体建立的免疫力可以抵抗一定剂量强毒的攻击。在质粒稳定性测定中,第9代次及以后的3个代次,20个菌落的VP2目的基因PCR检测结果均为阴性,表明该质粒在禽多杀性巴氏杆菌弱毒株G190E40内是不稳定的。
文摘Free fatty acids(FFAs) provide an important energy source and also act as signaling molecules.Medium to long-chain free fatty acids can activate the intracellular signal pathways in the pancreatic β-cells and play a role in regulating insulin secretion as an extracellular signal molecular via binding to the FFA receptor G protein-coupled receptor 40(GPR40). Furthermore,GPR40 is associated with several biological effects including cell proliferation and antiapoptosis of nerve cells.GPR40 act an important role in the connection of obesity and diabetes or cancers.GPR40 will probably become a novel kind of antidiabetic and anticancer drugs.
