共查询到20条相似文献，搜索用时 31 毫秒
1.
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
2.
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
3.
Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1×106ml-1, the percentages of conidium germination and appressorium formation were (97.98±0.67)% and (97.88±0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 pg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection. 相似文献
4.
Promoter trapping in Magnaporthe grisea 总被引：1，自引：0，他引：1
INTRODUCTION Magnaporthe grisea is a filamentous ascomy- cete that parasitizes economically important crops, such as barley, wheat, and rice (Talbot, 2003). It is also a good experimental model for studying fungal pathogenesis by both classical and molecular genetics (Dean, 1997). In recent years, many techniques have been developed to identify functional genes in M. grisea (Kamakura et al., 1999; Rauyaree et al., 2001; Takano et al., 2003; Irie et al., 2003; Lu et al., 2005). Of these… 相似文献
5.
Tong-bao LIU Guo-qing CHEN Hang MIN Fuc-heng LIN 《Journal of Zhejiang University. Science. B》2009,10(6)
Fasciclin family proteins have been identified as cell adhesion molecules in various organisms. In this study, a novel Magnaporthe oryzae fasciclin-like protein encoding gene, named MoFLP1, was isolated from a subtractive suppressive cDNA library and functionally analyzed. Sequence analysis showed that the MoFLP1 gene contains an open reading frame (ORF) of 1050 nucleotides encoding 349 amino acids with a calculated molecular weight of 35.85 kDa and a pl of 7.76. The deduced MoFLP1 protein contains a 17-amino acid secretion signal sequence and an 18-amino acid sequence with the characteristics of a glycosylphosphotidylinositol (GPI) anchor additional signal at its N- and C-terminuses, respectively. Potential N-glycosylation sites and domains involving cell adhesion were also identified in MoFLP1. Sequence analysis and subcellular localization by the expression of MoFLP1-GFP fusion construct in M. oryzae indicated that the MoFLP1 protein is probably localized on the vacuole membrane. Two MoFLP1 null mutants generated by targeted gene disruption exhibited marked reduction ofconidiation, conidiai adhesion, appressorium turgot, and pathogenicity. Our results indicate that fasciclin proteins play important roles in fungal development and pathogenicity in M. oryzae. 相似文献
6.
To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosomajaponicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5α. The amplified library contained more than 400 positive bacterial clones, which were then hybridized with forward and backward subtracted probes for differential screening, One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis, Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis. 相似文献
7.
Tong-bao Liu Guo-qing Chen Hang Min Fu-cheng Lin 《Journal of Zhejiang University. Science. B》2009,10(6):434-444
Fasciclin family proteins have been identified as cell adhesion molecules in various organisms. In this study, a novel Magnaporthe oryzae fasciclin-like protein encoding gene, named MoFLP1, was isolated from a subtractive suppressive cDNA library and functionally analyzed. Sequence analysis showed that the MoFLP1 gene contains an open reading frame (ORF) of 1050 nucleotides encoding 349 amino acids with a calculated molecular weight
of 35.85 kDa and a pI of 7.76. The deduced MoFLP1 protein contains a 17-amino acid secretion signal sequence and an 18-amino
acid sequence with the characteristics of a glycosylphosphotidylinositol (GPI) anchor additional signal at its N- and C-terminuses,
respectively. Potential N-glycosylation sites and domains involving cell adhesion were also identified in MoFLP1. Sequence analysis and subcellular
localization by the expression of MoFLP1-GFP fusion construct in M. oryzae indicated that the MoFLP1 protein is probably localized on the vacuole membrane. Two MoFLP1 null mutants generated by targeted gene disruption exhibited marked reduction of conidiation, conidial adhesion, appressorium
turgor, and pathogenicity. Our results indicate that fasciclin proteins play important roles in fungal development and pathogenicity
in M. oryzae.
Project supported by the National Natural Science Foundation of China (No. 30870101) and the Public Welfare Profession (Agriculture)
Research Project (No. 200803008), China 相似文献
8.
孙亮先 《泉州师范学院学报》2006,24(2):83-88
基因表达系列分析(Serial analysis of gene expression,SAGE)是功能基因组学研究中的一项功能强大的工具.对水稻幼苗SAGE文库的构建进行了探索试验,其主要操作步骤为:以磁珠法提取PolyA mRNA用于合成双链cDNA,再经过一系列的酶切、加接头序列、连接、PCR扩增,获得到26-bp的水稻双标签体,再将26-bp Ditag连接成标签链接体,并将其克隆进pZero-1载体中用于测序.DNA测序结果证明获得了水稻的SAGE标签.试验的各个中间步骤的结果均经过PCR验证.yh 相似文献
9.
Sequence analysis and expression pattern of MGTA1 gene in rice blast pathogen Magnaporthe grisea 总被引：1，自引：0，他引：1
INTRODUCTION Fungi cause most plant diseases of all groups of microbes. During their infection cycles, fungal pathogens must undergo two key processes: first, penetration through cuticles into host plant cells; second, colonization in host cells utilizing nutrients from their hosts. To penetrate host cells, fungi de-velop a series of specialized infection structures such as appressorium, penetration peg, and infection hypha. The appressorium-mediated penetration is a process typical of s… 相似文献
10.
谷胱甘肽过氧化酶2(GPx2),是生物体内重要的一类抗氧化酶,能够通过酶解过氧化氢阻断活性氧(ROS)的有害作用。本研究从七鳃鳗的血液c DNA文库中克隆了GPx2 c DNA全长和基因组DNA序列。序列分析表明,其c DNA全长为868 bp(Gen Bank序列号KM211710),包括53bp 5′非编码区、251bp 3′非编码区和564 bp开放阅读框,编码由187个氨基酸组成的多肽。基因组DNA序列(Gen Bank序列号KM211711)揭示了基因组特征含有2个外显子和1个内含子结构。系统发育分析表明,七鳃鳗GPx2(Lj GPx2)与其它的GPx2具有很高的同源性。利用实时荧光定量PCR检测到GPx2基因在七鳃鳗各组织中广泛表达,其中在口腔腺、心脏、卵、生殖腺中表达量较高。此外,用脂多糖(LPS)体内刺激七鳃鳗后发现,Lj GPx2 m RNA在生殖腺中表达量显著升高。本研究为探讨GPx2在七鳃鳗的免疫应答中的作用提供了理论依据。 相似文献
11.
Magnaporthe oryzae has been used as a primary model organism for investigating fungus-plant interaction. Many researches focused on molecular
mechanisms of appressorium formation to restrain this fungal pathogen. Autophagy is a very high conserved process in eukaryotic
cells. Recently, autophagy has been considered as a key process in development and differentiation in M. oryzae. In this report, we present and discuss the current state of our knowledge on gene expression in appressorium formation and
the progress in autophagy of rice blast fungi.
Project supported by the National Natural Science Foundation of China (Nos. 30671351 and 30870101) and the Hi-Tech Research
and Development Program (863) of China (No. 2002AA245041) 相似文献
12.
13.
Magnaporthe oryzae has been used as a primary model organism for investigating fungus-plant interaction. Many researches focused on molecular mechanisms of appressorium formation to restrain this fungal pathogen. Autophagy is a very high conserved process in eukaryotic cells. Recently, autophagy has been considered as a key process in development and differentiation in M. oryzae. In this report, we present and discuss the current state of our knowledge on gene expression in appressorium formation and the progress in autophagy of rice blast fungi. 相似文献
14.
Molecular analysis of microbial community in a groundwater sample polluted by landfill leachate and seawater 总被引：1，自引：0，他引：1
INTRODUCTION In some coastal cities of China, municipal solidwaste (MSW) is ultimately landfilled at seashorezones with cheaper land-value and less disturbance tonearby residents. These seashore landfills wereusually constructed without appropriate liners toprevent percolation of leachate into underlyingaquifers. Dramatic hydrochemical changes in theseaquifers have been detected as a result of the highorganic load of leachate. Furthermore, these aquiferswere inevitably infiltrated by sea… 相似文献
15.
Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B 总被引：3，自引：0，他引：3
Chen XH Chen Z Yao HP Chen F Zhu HH Zhou HJ 《Journal of Zhejiang University. Science. B》2005,6(4):288-294
INTRODUCTION Chronic hepatitis B virus (HBV) infection is aserious clinical problem because of its wide distribu-tion and possible adverse consequences, such as he-patic decompensation, cirrhosis and/or primary livercancer (PLC). The natural course of chronic HBVinfection is characterized by a series of hepatitic flaresor exacerbations and remissions (Ganem and Prince,2004). The severity, extent, duration and frequency ofhepatic histopathological changes in hepatitic flaresare d… 相似文献
16.
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
17.
抑制差减杂交(Suppression Subtractive Hybridization,SSH)是一种高效鉴定和分离克隆差异表达基因的新技术。目前,该技术在分子生物学研究的各个领域得到了广泛的应用。本对SSH的技术原理、差减cDNA库构建的技术流程、常见问题分析及优缺点等作了较全面的介绍,可为研究们提供参考。 相似文献
18.
毛赣鸣 《河南科技学院学报》2014,(11):88-92
阐述了世界图书馆立法史的历程和立法理念的三个阶段,大致可分为划分为：图书馆立法早期;图书馆立法普及期;图书馆网络化立法阶段。重点表述了当代国外图书馆法案动向和国际图联立场,针对图书馆法权和版权关系进行了简要分析。 相似文献
19.
20.
利用西方蜜蜂雄蜂头部5’LongSAGE文库中的一组标签序列,在蜜蜂基因组序列第9号连锁群上定位一个新的西方蜜蜂表皮蛋白基因转录起始位点,并克隆了该基因的eDNA序列,继而利用此cDNA序列分析了该基因的内含子和外显子结构．分析结果显示：该基因的DNA序列长1253bp,含有4个外显子和3个内含子,其cDNA长598bp,编码一个169AA的富含Val和Pro残基（23．67％,20．12％）多肽序列．BLAST分析发现,该蛋白与已知的4个昆虫表皮蛋白序列的相似性为47．54％,且其C末端有一个共有基序,说明这5个蛋白可能属于同一个表皮蛋白家族．该研究结果提供了一个新的蜜蜂表皮蛋白基因,且该基因在雄蜂头部表达水平较高． 相似文献