TargetedGeneReplacementinFungalPathogensviaAgrobacterium...

Genome sequence data on fungal pathogens provide the opportunity to carry out a reverse genetics approach to uncover gene function. Efficient methods for targeted genome modifications such as knockout and in locus over-expression are in high demand. Here we describe two efficient single-step cloning strategies for construction of vectors for Agrobacterium tumefaciens -mediated transformation (ATMT). Targete......阅读全文

Targeted-Gene-Replacement-in-Fungal-Pathogens-via-Agrobacterium-...

Genome sequence data on fungal pathogens provide the opportunity to carry out a reverse genetics approach to uncover gene function. Efficient meth

Targeted-Gene-Replacement-in-Fungi-Using-a-SplitMarker-Approach

Targeted gene replacement is one of the primary strategies for functional characterization of fungal genes and several methods have been developed

基因转型(gene-transformation)

目的带有特定基因的质体在分子生物研究上,是一个很重要的工具,将质体送入细菌的过程称为基因转形,经由基因转型可使质体在细菌中大量复制,以备进一步研究。本实验将把你在前面转殖实验中cDNA和质体DNA的连结反应送入细菌。 你将从本实验学习如何进行基因转型作用。原理早在1970年左右,有人发现细菌经由冰冷

Use-of-the-GUS-Reporter-Gene

One of the most important considerations in the expression of heterologous proteins in plants is the choice of promoter. The study of promoter act

Gene-Structure-Annotation-at-PlantGDB

The accurate identification of exons and introns that comprise a complete plant gene structure can be a time-consuming and challenging task. Novel

FOSB-gene-expression-and-drug-abuse

Drug addiction is associated with long-term behavioral changes, suggesting a long-lived transcriptional regulator that responds to chronic drug exposu

Reverse-Transfection-for-Gene-Function-Analysis

This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured

科学家利用基因组编辑家蚕大量表达蜘蛛丝

  近日,中国科学院分子植物科学卓越创新中心/植物生理生态研究所谭安江研究组利用基因定点替换的方法在家蚕丝腺和蚕茧中大量表达蜘蛛丝蛋白。   蜘蛛丝是自然界中机械性能最好的天然蛋白纤维,其强度甚至高于用于制作防弹衣的凯夫拉纤维,在工业、医疗和国防中有广泛应用前景。如何大量获取蜘蛛丝纤维是一直以来难

科学家利用基因组编辑家蚕大量表达蜘蛛丝

  近日,中国科学院分子植物科学卓越创新中心/植物生理生态研究所谭安江研究组利用基因定点替换的方法在家蚕丝腺和蚕茧中大量表达蜘蛛丝蛋白。   蜘蛛丝是自然界中机械性能最好的天然蛋白纤维,其强度甚至高于用于制作防弹衣的凯夫拉纤维,在工业、医疗和国防中有广泛应用前景。如何大量获取蜘蛛丝纤维是一直以来难

基因敲除技术概述(四)

[13]。2.3.2 RNAi基因敲除的优点及应用①.比用同源重组法更加简便,周期大大缩短。②.对于哺乳动物,如对于一些敲除后小鼠在胚胎时就会死亡的基因,可以在体外培养的细胞中利用RNAi技术研究它的功能。③.由于RNAi能高效特异的阻断基因的表达,它成为研究信号传导通路的良好工具。④.RNAi还被

差异基因表达研究方法介绍(DDPCR;GENEFISHING;GENE-CHIP)

差异基因表达的研究受到了广泛的关注,常用的技术有DD-PCR;GENE-FISHING;GENE CHIP等。简单介绍如下: DDRT -PCR技术即mRNA差异显示聚合酶链式反应技术,此技术是以PCR技术和聚丙烯凝胶电泳技术为基础,结合银染或放射性自显影等显色技术,能快速有效地

PNAS:利用基因组编辑家蚕大量表达蜘蛛丝

  蜘蛛丝是自然界中机械性能最好的天然蛋白纤维,其强度甚至高于用于制作防弹衣的凯夫拉纤维,在工业、医疗和国防上都有着广泛的应用前景。但是如何大量获取蜘蛛丝纤维是一直以来难以解决的问题。  来自中科院分子植物科学卓越创新中心/植物生理生态研究所谭安江研究组题为“Mass spider silk pro

PCR-Primers-For-Gene-Expression-Detection-or-Quantification

Why PrimerBank?PrimerBank is a public resource for PCR primers. These primers are designed for gene expression detection or quantification (real-time

Double-Stranded-RNA-Induced-Gene-Expression

One defense against viral infection is provided by PKR, double-stranded RNA activated protein kinase. When PKR interacts with dsRNA found in cells dur

Yeast-Gene-knockout-using-Oligo/PCR

Universal primers for gene knock-out using dominant drug markers: Kan, Clonat, and Hygromisin-B. Forward primer: 5’ TCAGGGGCATGATGTGACT 3’Reverse prim

Control-of-Gene-Expression-by-Vitamin-D-Receptor

The vitamin D receptor, VDR is the mediator of all genomic actions of vitamin D3 and its analogs. It belongs to a family of ligand induced transcripti

转基因——基因标靶

Gene Targeting Outline (University of Michigan Transgenic Animal Model Core)This is a brief outline of the steps necessary to produce mice with a muta

Functional-Genomics-and-Structural-Biology-in-the-Definition-of-Gene...

By mid-2007, the three-dimensional (3D) structures of some 45,000 proteins have been solved, over a period where the linear structures of millions

基因芯片(gene-chip)的原理

基因芯片(gene chip)的原型是80年代中期提出的。基因芯片的测序原理是杂交测序方法,即通过与一组已知序列的核酸探针杂交进行核酸序列测定的方法,在一块基片表面固定了序列已知的八核苷酸的探针。当溶液中带有荧光标记的核酸序列TATGCAATCTAG,与基因芯片上对应位置的核酸探针产生互补匹配时,通

报告基因(report-gene)的应用

报告基因被广泛地应用于细胞生物学中的基因表达和细胞相关内容的研究。常用的报告基因主要有GUS基因、CAT基因、hGH基因、Cato2ase基因、绿色荧光蛋白基因及萤火虫荧光素酶基因等。在这些报告基因中萤火虫荧光素酶基因因其表达产物-荧光素酶敏感性高,测定方法快速且宜于掌握,线性好,并且荧光素酶产生的

GENEπ数字PCR技术应用教程

数字 PCR( digital PCR ,dPCR ) 下一代DNA/RNA扩增技术。原理是将一个PCR 反应体系分配到大量微小的反应单元中,在每个微反应器中包含或不包含 1 个或多个拷贝的目标核酸分子 (DNA 模板) ,进行“单分子模板”PCR 扩增。扩增结束后,通过阳性反应单元( 通过

Overview-of-telomerase-protein-component-gene-hTert-Transcriptional

Telomerase is an enzyme which replicates the terminal sequences of eukaryotic chromosomes, namely the telomeres. Cells which have an unlimited replica

RNAi片段siRNA设计原则

RNAi target selection rules:Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG).S

基因治疗按基因操作分类介绍

一类为基因修正(gene correction)和基因置换(gene replacement),即将缺陷基因的异常序列进行矫正,对缺陷基因精确地原位修复,不涉及基因组的其他任何改变。通过同源重组(homologous recombination)即基因打靶(gene targetting)技术将外源

关于基因治疗的按基因操作介绍

  一类为基因修正(gene correction)和基因置换(gene replacement),即将缺陷基因的异常序列进行矫正,对缺陷基因精确地原位修复,不涉及基因组的其他任何改变。通过同源重组(homologous recombination)即基因打靶(gene targetting)技术将

基因治疗的操作分类

一类为基因修正(gene correction)和基因置换(gene replacement),即将缺陷基因的异常序列进行矫正,对缺陷基因精确地原位修复,不涉及基因组的其他任何改变。通过同源重组(homologous recombination)即基因打靶(gene targetting)技术将外源

基因治疗按基因操作分类介绍

  一类为基因修正(gene correction)和基因置换(gene replacement),即将缺陷基因的异常序列进行矫正,对缺陷基因精确地原位修复,不涉及基因组的其他任何改变。通过同源重组(homologous recombination)即基因打靶(gene targetting)技术将

Lissencephaly-gene-(LIS1)-in-neuronal-migration-and-development

Integration of pathways that regulate nucleokinesis during neuronal migration and a model of LIS1 mediating CLIP-170 interactions with the dynein/dyna

Gene-splicing-and-mutagenesis-by-PCRdriven-overlap-extension

实验概要        Extension of overlapping gene segments by PCR is a simple, versatile technique for site-directed mutagenesis and gene splicing.Initial

基因表达轮廓(gene-expressed-profile)技术2

经过这轮RDA过程,Tester中的目的DNA将得到第一次富集。将第一轮产物更换新接头,进行第二轮RDA过程,目的DNA可得到进一步的富集。该技术假阳性很低。但仍不能解决个体mRNA在丰度上存在巨大差异的问题,当靶序列浓度较低时,其富集受抑制。3:抑制消减杂交(suppression subtrac