农杆菌介导的蝴蝶兰基因转化系统的建立   总被引：9，自引：1，他引：9  

柴明良  金斗焕 《园艺学报》2004,31(4):537-540

 针刺后的蝴蝶兰‘White Hikaru’的类原球茎(PLB)，与含绿色荧光蛋白基因( )和潮霉素磷酸转移酶基因(^p￡)的pCAMBIA 1300一SmGFP的根瘤农杆菌LBA4404 共培养，培育出了转基因的蝴蝶兰。经绿色荧光蛋白检测和Southern印迹，证实了再生植株中含cop基因和hpt基因。  相似文献   

棉铃虫核型多角体病毒分子生物学和基因工程研究进展   总被引：1，自引：1，他引：1  

刘艳荷  张传溪 《植物保护学报》2004,31(3):320-328

棉铃虫核型多角体病毒(HaSNPV)是棉铃虫专一性病原物,隶属于杆状病毒科核型多角体病毒属.对其分子生物学和基因工程的研究主要包括以下几个方面:测定了HaSNPV G4株和C1株基因组核苷酸全序列,并与其它病毒进行了同源性比较;研究了HaSNPV部分基因的结构、转录、表达及其功能.构建了HaSNPV Bac to Bac杆状病毒表达系统;重组病毒杀虫剂的研究为HaSNPV大面积防治棉铃虫展示了广阔的前景.随着棉铃虫核型多角体病毒分子生物学和基因工程研究的不断深入,重组病毒杀虫剂将在棉铃虫综合防治中发挥更为重要的作用.  相似文献   

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   

果树农艺性状基因的分子标记及其应用   总被引：2，自引：1，他引：2  

乔玉山  房经贵  沈志军  章镇 《果树学报》2004,21(2):158-163

总结了果树农艺性状基因标记策略和已获得的分子标记及其在果树上的应用。集团混合分离分析法(BSA)、平行芽变分析法(PSA)和已知信息捕捉法(KIH)是果树农艺性状基因标记筛选的主要方法。目前已获得了大量果树农艺性状基因的分子标记,这些标记是果树基因作图、图位克隆、分子辅助选择和种质资源鉴定等遗传研究与育种实践的有用工具。  相似文献   

以Taichung29为背景的小麦抗条锈病近等基因系转育进展   总被引：6，自引：0，他引：6  

徐世昌  吴立人  万安民  王凤乐  赵文生  牛永春 《植物保护》2004,30(2):19-22

自1988年起,系统开展了以Taichung29为背景的小麦抗条锈病近等基因系转育及其基础性研究。采用回交法和系谱法相结合的转育方法,以春性品种Taichung29为轮回亲本作母本,分别与25个抗性供体即中国小麦条锈病菌鉴别寄主和国际上重要的抗条锈基因载体品种杂交、回交和自交。通过基因推导分析、单体分析和SSR标记技术检测目的基因,选育抗条锈近等基因系,现已获得重要进展,成功选育出8个以Taichung29为背景的抗条锈病单基因近等基因系,即Taichung29*6／Yr1、Taichung29*6／Yr2、Taichung29*6／Yr5、Taichung29*6／Yr7、Taichung29*6/Yr9、Taichung29*6／Yr10、Taichung29*6/YrSpP、Taichung29*6/YrKy2。另有9个组合转育获得343个抗性稳定株系,正检测其目的基因,3个组合转至BC₆F₃,自交纯合筛选抗性稳定株系,5个组合转至BC₅,继续回交转育。  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   

Validation of the specificity and sensitivity of species-specific primers that provide a reliable molecular diagnostic for <Emphasis Type="Italic">Xiphinema diversicaudatum</Emphasis>, <Emphasis Type="Italic">X. index</Emphasis> and <Emphasis Type="Italic">X. vuittenezi</Emphasis>     

Judith Hübschen  Lilo Kling  Ulrike Ipach  Volker Zinkernagel  Nathalie Bosselut  Daniel Esmenjaud  Derek J.F. Brown  Roy Neilson 《European journal of plant pathology / European Foundation for Plant Pathology》2004,110(8):779-788

Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry.  相似文献   

蜱传性疾病--莱姆病螺旋体研究进展   总被引：6，自引：0，他引：6  

廖德丰  陈季武  谢萍  傅蓓蓓  朱振勤 《中国兽医寄生虫病》2004,12(2):36-39

莱姆病是一种蜱传性人兽共患病,在我国北方各省广泛流行,其病原体为伯氏疏螺旋体。莱姆病可使皮肤、心脏、关节和神经系统等多种组织器官受损。本文综述了莱姆病螺旋体的形态、种下分类、生化特征、传播媒介和贮存宿主等方面的研究进展"  相似文献   

鹅细小病毒VP2基因在大肠杆菌中的表达及纯化   总被引：2，自引：0，他引：2  

贺云霞  王君伟  王立群  Ulrich Neumann 《中国兽医学报》2004,24(2):126-128

将鹅细小病毒 (GPV) VP2基因插入原核表达载体 p PROEX- HTb,获得重组表达质粒 p PROEX- HTb- VP2。将其转化大肠杆菌 DH5α,用 IPTG诱导表达 ,表达菌体蛋白中可产生与预期大小相符的约 72 0 0 0的蛋白。以光密度扫描对该表达产物进行定量分析 ,表达产物约占菌体总蛋白的 14 %。经 His- tag金属螯合层析纯化 ,获得纯度较高的 6 His- VP2融合蛋白。 Western- blot结果表明 ,所得蛋白与鹅抗 GPV高免血清有较好的免疫反应性 ,说明在 VP2的 N端融合 6个组氨酸不影响其与特异抗体结合的活性。  相似文献