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LU Jian-ping LIU Tong-bao YU Xiao-yun LIN Fu-cheng 《Journal of Zhejiang University. Science. B》2005,(2)
A mature appressorium cDNA library of rice blast fungus, Magnaporthe grisea, was constructed in a ?TriplEx2 vector by SMARTTM cDNA library containing 2.37?106 independent clones about 100% of which harbor foreign cDNA inserts with average size of 660 bp. Of 9 randomly selected clones, 2 expressed sequence tags (ESTs) sequences did not have homologous EST sequences of M. grisea in GenBank. The appressorium cDNA library is suitable for gene expression analysis and function analysis of the late stages of appressorium formation and the early stages of penetration of M. grisea. 相似文献
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INTRODUCTION 1,25-dihydroxyvitamin D3[1,25(OH)2D3], the biologically active metabolite of vitamin D3, is a secosteroid hormone that not only regulates bone and calcium/phosphate metabolism but also regu-lates a number of other biological activities, in-cluding modulation of the immune response via specific receptors expressed in antigen presenting cells (APC) and activated T cells. Recently, in-creasing evidence showed that the modulatory role of 1,25(OH)2D3 on T cell phenotype and … 相似文献
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Jin QC Dong HT Peng YL Chen BS Shao J Deng Y Dai CE Fang YQ Lou YC Li YZ Li DB 《Journal of Zhejiang University. Science. B》2007,8(2):88-97
Appressorium is an infection structure of the phytopathogenic fungus Magnaporthe grisea. Analysis of gene expression profiles ofappressorium development provides insight into the molecular basis of pathogenicity and control of this fungal plant disease. A cDNA array representing 2927 unique genes based on a large EST （expressed sequence tag） database ofM. grisea strain Y34 was constructed and used to profile the gene expression patterns at mycelium and appressorium maturation stages. Compared with mycelia, 55 up-regulated and 22 down-regulated genes were identified in mature appressoria. Among 77 genes, 16 genes showed no similarity to the genome sequences of M. grisea. A novel homologue of peptidyl-prolyl cis-trans isomerase was found to be expressed at low-level in mature appressoria of M. grisea. The results indicated that the genes such as pyruvate carboxylase, phospholipid metabolism-related protein and glyceraldehyde 3-phosphate dehydrogenase involved in gluconeogenesis, lipid metabolism and glycolysis, showed differential expression in mature appressoria. Furthermore, genes such as PTHll, beta subunit of G protein and SGTI involved in cell signalling, were expressed differentially in mature appressoria. Northern blot analysis was used to confirm the cDNA array results. 相似文献
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5’LongSAGE标签得自于全长mRNA分子的5’末端的前19 nt.该研究利用定位在西方蜜蜂基因组中的一个预测基因座LOC724521的2条5’LongSAGE标签序列作为5’引物,通过RT-PCR克隆了该预测基因座的2条长335 bp和337 bp的cDNA序列（GenBank登录号：GU358205,GU358204）.此cDNA编码一条长88个氨基酸残基的多肽.用所克隆的cDNA序列对基因座LOC724521进行功能注释发现,该基因含有3个外显子和2个＂GT-AG＂型内含子.5’LongSAGE标签序列的基因组定位结果显示：基因座LOC724521在雄蜂的头部中表达丰度很高,RNA PolⅡ可从5个转录起始位点（TSS）上以不同效率起始转录,该基因的59%和31%的转录是从2个优势TSS上起始.有趣的是,有一条5’LongSAGE标签序列被定位在内含子区,暗示该基因存在外显子的可变性选择现象.该研究结果不仅在转录水平上证实了软件预测的基因座LOC724521确实存在,同时揭示了该基因存在可变性转录起始位点和可变性外显子选择等转录调控机制. 相似文献
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鹅MYL1基因的克隆及胚胎期表达特征分析 总被引：1，自引：0，他引：1
肌球蛋白是肌纤维的主要组成成分,在肌肉生长和收缩过程中具有重要作用.本研究以鹅（Anser anser）肌肉总RNA为模板,采用RACE的方法,克隆肌球蛋白轻链1（MYL1）基因全长cDNA序列,并进行生物信息学分析.运用实时荧光定量PCR检测MYL1基因在鹅胚胎期的表达特征.结果表明,鹅MYL1基因全长cDNA为1542 bp,编码193个氨基酸残基的肽链.预测鹅MYL1蛋白等电点5.12,分子量21.75 KD,具有典型的EFh和FRQ1结构域,且不同物种间EFh氨基酸序列高度同源.荧光定量PCR结果显示鹅胚胎期MYL1基因mRNA表达量总体呈现上升后下降的趋势,该基因在胚胎期E7就有表达,E7以后表达量逐渐上升,在E18表达量达到高峰后下降.研究结果首次提供了鹅肌肉组织主要结构基因MYL1的全序列和蛋白特征信息,并揭示该基因在鹅肌肉组织发生和发育的功能. 相似文献
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In this study the MTP1 gene, encoding a type III integral transmembrane protein, was isolated from the rice blast fungus Magnaporthe oryzae. The Mtp1 protein is 520 amino acids long and is comparable to the Ytp1 protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP (green fluorescent protein) fusion expression results indicate that Mtp1 is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily expressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for pathogenicity. The Deltamtp1 mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use. 相似文献
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2020年的诺贝尔生理学或医学奖授予了在发现丙肝病毒(HCV)方面作出突出贡献的哈维·阿尔特(Harvey J. Alter)、迈克尔·霍顿(Michael Houghton)和查尔斯·赖斯(Charles M. Rice)。然而,从1989年霍顿捕获HCV到1997年赖斯证明HCV能引发肝炎,间隔了8年时间。这看似一步之遥的距离何以耗时如此之久?文章通过回顾文献,对这一时期的研究脉络进行了梳理,明确了包括赖斯团队在内的不同研究者对于推进HCV相关认识所作的具体贡献。同时,针对研究过程中的两个关键节点——病毒RNA基因组3′末端的准确测序和感染性分子克隆的建立,文章从技术储备、路径依赖和目标定位的角度分析了赖斯团队为何能在科学研究竞争中拔得头筹。 相似文献
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An Inexpensive Gel Electrophoresis-Based Polymerase Chain Reaction Method for Quantifying mRNA Levels 
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下载免费PDF全文 William D. Bradford Laty Cahoon Sara R. Freel Laura L. Mays Hoopes Todd T. Eckdahl 《CBE life sciences education》2005,4(2):157-168
In order to engage their students in a core methodology of the new genomics era, an ever-increasing number of faculty at primarily undergraduate institutions are gaining access to microarray technology. Their students are conducting successful microarray experiments designed to address a variety of interesting questions. A next step in these teaching and research laboratory projects is often validation of the microarray data for individual selected genes. In the research community, this usually involves the use of real-time polymerase chain reaction (PCR), a technology that requires instrumentation and reagents that are prohibitively expensive for most undergraduate institutions. The results of a survey of faculty teaching undergraduates in classroom and research settings indicate a clear need for an alternative approach. We sought to develop an inexpensive and student-friendly gel electrophoresis-based PCR method for quantifying messenger RNA (mRNA) levels using undergraduate researchers as models for students in teaching and research laboratories. We compared the results for three selected genes measured by microarray analysis, real-time PCR, and the gel electrophoresis-based method. The data support the use of the gel electrophoresis-based method as an inexpensive, convenient, yet reliable alternative for quantifying mRNA levels in undergraduate laboratories. 相似文献