LUYOR-3415一区文献汇总

仪橙科技实验室产品系列涵盖激发光源、紫外交联仪、紫外灯、细胞光毒性  辐照仪、光反应仪等，广泛应用于农学、植物学、生物学、化学、医学、 材料学等不同领域的科学研究。

今天整理几篇，希望帮助正在为实验发愁的你们哦

期刊名称：《Nature》(IF=48.5,   中科院一区top 期刊)

实验方法：

R.solanacearum spot inoculations and quantification of bacterial populations

R.solanacearum spot inoculation assay and statistics of R.solanacearum populations were

performed as previously reported.R.solanacearum was grown in phi medium,washed and

resuspended in sterile water.OD600 was adjusted to 0.02 for inoculation.M.truncatula plants

were cul tivated in BNM medium for 4-5 days.Sterile needles were used to pierce the root

segments(2cm from the root tip)of each plant for co-inoculating Nod factor with Ralstonia or H2O with Ralstonia viafilter paper.To determine R.solanacearum populations 5 dpi in the inoculation    area of roots,root segments were individually placed in tubes and ground.The material was

resuspended in sterile waterand serial dilutions were plated on Phi medium.Colonies were counted after incubation at 28℃ for 48h,with eGFP colonies counted under a  fluorescent protein lamp （LUYOR-3415)

如先前报道所述，进行了R. solanacearum斑点接种实验及细菌种群定量分析，并统计了R. solanacearum种群数量。R. solanacearum在phi培养基中培养后，经洗涤并重悬于无菌水中；接种时将OD600值调整至0.02。Truncatula植株在 BNM 培养基中培养4-5天。使用无菌针头刺穿每株植物根段（距根尖2厘米处），通过滤纸同时接种Nod因子与Ralstonia菌或水与Ralstonia菌。为测定接种后第5天根部接种区域的R. solanacearum种群数量，将各根段单独置于试管中研磨，将材料重悬于无菌水中，进行系列稀释后涂布于Phi培养基上。于28℃培养48小时后计数菌落，其中eGFP阳性菌落使用荧光蛋白灯进行计数。（非原作者翻译）

实验方法

期刊名称：《Molecular Plant》(IF=27.5,中科院一区top期刊)

Protein expression,extraction,and purification

The BL21(DE3)strain was used to express GST-or His-tagged fusion proteins in E.coli,and

GFP-tagged fusion proteins were expressed in N.benthamiana.For protein expression in BL21,an overnight culture of BL21 harboring corresponding expression constructs was diluted in 200 ml LB medium.The culture was grown at 37℃ to an OD600 of 0.6-0.8,supplemented with 1 mM IPTG,  and shaken for another 3h.Cells were harvested by centrifugation at 4000 g and 4℃ for 15 min.   For protein expression in tobacco,the GV3101 strain harboring the corresponding expression

constructs was transfected into tobacco leaves as described previously.After 1-3 days of culture,  fluorescent areas detected with a portable GFP excitation light source were cut out and treated as indicated.

蛋白质表达、提取与纯化：采用BL21(DE3)菌株在大肠杆菌中表达GST或His标签融合蛋白，而GFP标签融合蛋白则在本氏烟草中表达。对于BL21菌株的蛋白质表达实验，将携带相应表达载体的BL21菌液过夜培养后，用200毫升LB培养基稀释。培养物在37℃条件下培养至OD600值达0.6-0.8，加入1 mM IPTG 后继续振荡培养3小时。随后通过4000 g、4℃离心15分钟收集细胞。对于烟草中的蛋白质表达实验，将携带相应表达载体的GV3101菌株按先前方法转染至烟草叶片。培养1-3天后，使用便携式 GFP 激发光源检测荧光区域，并按指定方法进行处理。（非原作者翻译）