苏云金芽胞杆菌防治植物寄生线虫的研究进展   总被引：2，自引：0，他引：2  

余子全  周燚  孙明  喻子牛 《植物保护学报》2004,31(4):418-424

植物寄生线虫是一类重要的植物病原生物,给农业生产造成巨大的损失.传统方法防治植物寄生线虫都存在一定的局限性.研究发现苏云金芽胞杆菌对线虫有毒杀活性,为防治植物寄生线虫寻找到了一条新途径.文章介绍了苏云金芽胞杆菌防治植物寄生线虫的研究现状,着重阐述了苏云金芽胞杆菌的杀线虫晶体蛋白及其基因、杀线虫机制等方面的研究进展,并探讨了其中存在的问题及其对策.  相似文献   

芽孢杆菌绿色荧光蛋白标记及其在小麦体表定殖的初探   总被引：17，自引：0，他引：17  

田涛  亓雪晨  王琦  梅汝鸿 《植物病理学报》2004,34(4):346-351

 将来自质粒pAD4412的启动子和绿色荧光蛋白基因gfpmut3a插入大肠杆菌-枯草芽孢杆菌穿梭载体pBE2,构建成芽孢杆菌表达载体pGF P4412,用其转化野生型生防芽孢杆菌83-6和A-47等8个菌株,均得到良好的发光表型。质粒稳定性实验表明重组质粒pG FP4412稳定性为92%。借助荧光显微镜对gfp标记的菌株A-47-gfp在小麦体表的定殖进行初步的研究。结果表明:A-47-gfp能够在小麦根际及小麦体表定殖(包括根表和茎叶表面);相对于在茎叶表面定殖的A-47-gf p在根表定殖的菌体与根的结合更为牢固;从根基到根尖A-47-gfp的定殖量有明显的减少趋势。  相似文献   

不同地域环境对枸杞蛋白质和药用氨基酸含量的影响   总被引：12，自引：0，他引：12  

张晓煜  刘静  袁海燕  亢艳莉  张映琪 《干旱地区农业研究》2004,22(3):100-104

根据2000～2001年我国北方六省(区)多点枸杞采样资料和田间试验资料,利用K-均值聚类方法和非线性回归方法,对比分析了宁杞1号枸杞蛋白质和9种药用氨基酸含量的差异和特点,以及生态因子对它们的影响。分析结果表明,不同地域栽种的宁杞1号枸杞蛋白质和9种药用氨基酸含量的变异系数分别为16.58%和16.22%,除品种因子的决定作用以外,环境条件对枸杞蛋白质含量有一定作用,其中土壤水解氮含量对蛋白质和氨基酸合成有一定作用,二者呈对数关系。  相似文献   

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes     

JIANG Xun  ZENG Yao-ying  HE Xian-hui  XU Li-hui  DI Jing-fang  FENG Zheng  ZHAO Jing-xian  WANG Qing  WANG Tong  SHI Jian-bo 《园艺学报》2004,20(6):924-928

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.  相似文献   

Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion     

ZHANG Liang  WANG Wei  RUAN Xu-zhong 《园艺学报》2004,20(9):1553-1556

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.  相似文献   

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver     

LIU Ping-ping  ZHANG Yuan 《园艺学报》2004,20(11):2053-2057

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.  相似文献   

Changes of MAPKs expression in rat hippocampal neurons after sleep deprivation     

ZHANG Qian  CHENG Jiang-tao  WANG Shu-chun  QIAO Peng  WANG Yi-ling  WU Jing-lan  WANG Yu-ruo 《园艺学报》2004,20(10):1874-1877

AIM: To explore the change and the possible role of MAPKs in rat hippocampus neuron after sleep deprivation. METHODS: The morphology of hippocampus neuron after sleep derivation was observed by TUNEL and HE staining, the activity of ERK was assayed by β-liquid scintillation counting and the expression of JNK was detected by Western blot. RESULTS: In paradoxical sleep deprivation (PSD) group, the number of apoptotic cells in hippocampus was increased. The scores of ERK activity were 1 764.00±941.56. Compared with control groups, the ERK activity was obviously decreased (P<0.05). The JNK expression was 87.5%, which was higher than that in control group. CONCLUSION: These results provide some important evidences that the sleep deprivation could cause changes in MAPKs activity, which may be related to the mechanism of hippocampus neuron apoptosis.  相似文献   

家鸡Leptin成熟肽cDNA的克隆、重组蛋白表达及纯化   总被引：4，自引：0，他引：4  

施振旦  邵西兵  方梅霞  于迎春  刘颖  陈楠 《中国兽医学报》2004,24(3):258-260

从 18周龄鸡卵巢组织中抽提总 RNA,使用六聚体随机引物反转录后 ,用鸡 L eptin(瘦素 )特异性引物扩增出鸡L eptin编码区第 5 2～ 4 6 0 bp的长度为 4 0 9bp的 c DNA片段。根据鸡 L eptin的 3′端第 4 6 0～ 4 92 bp序列设计了 3条部分相互重叠并且顺序串联延伸的下游反义引物 ,并在最后 1条引物 3′端连接上 Eco R 切点 ;在以上用于反转录扩增的 5′端引物的 5′端连接一 Bam H 切点。用该 5′端引物分别与 3个 3′端引物配对 ,利用反转录扩增出的 4 0 9bp的L eptin c DNA片段作为第 1模板进行扩增 ,扩增产物再作为模板与下一引物对再次扩增。经 3次扩增得到编码鸡 L ep-tin成熟肽的全长 4 5 1bp的 c DNA序列。将该 4 5 1bp的 L eptin c DNA序列经 Bam H 和 Eco R 双酶切后 ,克隆入表达质粒 p RSET A的 Bam H 和 Eco R 两酶切位点之间 ,构建成表达质粒 p L ep- SCAU。转化有重组表达质粒 p L ep-SCAU的大肠杆菌 BL 2 1(DE3)在 L B培养基中培养后 ,经 IPTG诱导表达出相对分子质量为 2 0 10 0的鸡 L eptin融合蛋白和少量 4 0 2 0 0的 L eptin融合蛋白。L eptin融合蛋白的表达在 IPTG浓度为 0 .0 5 mmol/ L 时达到最高 ,占总菌体蛋白的 32 .6 %。用 Ni- NTA凝胶从 7L 发酵培养菌裂解液中纯化出 180 m g左右  相似文献   

禽大肠杆菌外膜蛋白基因C(ompC)的序列分析   总被引：1，自引：0，他引：1  

曹素芳  黄青云  张煜坤  温纳相  陈红英 《中国兽医学报》2004,24(4):332-334

根据 Gen Bank中人源大肠杆菌 K- 12外膜蛋白基因 C(omp C)的核苷酸序列设计引物 ,应用 PCR方法从禽大肠杆菌 O2 、O78株及它们的融合双价弱毒菌株 O2 ,78(Norr Chlr)中分别扩增得到 omp C基因 ,序列测定及分析比较发现 ,3个菌株的 om p C基因均由 170 2 nt组成 ,核苷酸序列完全相同 ,只有 1个大的开放性阅读框 (ORF) ,长 10 92 bp,编码由 36 3个氨基酸组成的前 Om p C蛋白 ,前 2 1个氨基酸残基组成信号肽 ,成熟的 Omp C蛋白由 342个氨基酸残基组成 ,Mr为 4 0 0 0 0。其氨基酸序列也完全相同。从基因水平上证明了禽大肠杆菌 O2 、O78株及融合双价弱毒菌株 O2 ,78(NorrChlr)存在相同的外膜蛋白 C抗原 ,从而为进一步研究 Omp C蛋白的免疫原性奠定了基础  相似文献   

融合表达淋巴细胞脉络丛脑膜炎病毒和卵清白蛋白CD8+T细胞表位的减毒鼠伤寒沙门氏菌的构建与鉴定   总被引：3，自引：0，他引：3  

王芳  焦新安  潘志明  张晓明  黄金林  Richard Lo-Man  Claude Leclerc  刘秀梵 《中国预防兽医学报》2004,26(1):14-17

依据淋巴细胞脉络丛脑膜炎病毒(LCMV)主要保护性抗原CD8 T细胞表位VRRPQASGVYMGNLTAQ和卵清白蛋白(OVA)CD8 T细胞表位SIINFEKL,设计、合成两条编码LCMV和OVA CD8 T细胞表位的寡核苷酸片段,经退火后,克隆入绿色荧光蛋白表达质粒pYAGFP,经PCR扩增和序列测定分析,证实成功构建T细胞表位与绿色荧光蛋白融合表达的重组质粒pYAGFPL-O,将此重组质粒pYAGFPL-O转化减毒鼠伤寒沙门氏菌X4550,获得重组沙门氏菌X4550(pYAGFPL-O).用SDS-PAGE电泳测得重组菌表达的融合蛋白约为30kD.同时,荧光显微镜下观察到X4550(pYAGFPL-O)发出黄绿色荧光.这些结果表明LCMV和OVA T细胞表位已成功表达.重组菌X4550(pYAGFPL-O)的获得为研究沙门氏菌载体携带外源抗原的T细胞应答规律及调控机理打下了重要基础.  相似文献