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1.
A mature appressorium cDNA library of rice blast fungus, Magnaporthe grisea, was constructed in a λTriplEx2 vector by SMART?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 相似文献
2.
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
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.
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
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.
孙亮先 《泉州师范学院学报》2004,22(2):94-99
对GenBank中9215条来源于水稻(Oryza sativa L.ssp．Indica)胚乳cDNA库的3’EST进行了分析，获得20个淀粉合成相关基因的表达丰度信息，研究发现淀粉合成的5个关键酶的编码基因，ADPG焦磷酸化酶基因、ADP—葡萄糖淀粉合成酶、淀粉分支酶、淀粉去分支酶、蜡质基因的表达丰度极高，表明该时期水稻胚乳内淀粉合成反应非常强烈，研究发现在淀粉合成途径中存在功能相关的基因协同表达的现象，章结合所获得的基因表达信息对淀粉合成的分子机理进行了探讨。 相似文献
7.
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
8.
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
9.
1IntroductionNeural stemcells(NSCs)are a subtype of progenitorcells in the nervous systemthat can differentiate intoneurons and glia[1-3].Due to their feature of self-re-newal,NSCs have expectations for treatment of ner-vous system diseases such as Parkin… 相似文献
10.
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
11.
Xin-chun Zhang Xin Yu Hui-juan Zhang Feng-ming Song 《Journal of Zhejiang University. Science. B》2009,10(10):731-739
We cloned and characterized a rice gene OsBIABPI encoding an AMP-binding protein. The full-length cDNA of OsBIABP1 is 1912-bp long and is predicted to encode a 558-aa protein. OsBIABP1 contains a typical AMP-binding signature motif and shows high similarity to members of AMP-binding protein family. OsBIABP1 is expressed in stems, leaves and flowers of rice plants, but is not expressed, or expressed at a very low level, in rice roots. The expression of OsBIABP1 was induced by some defense-related signal molecules, e.g., salicylic acid (SA), benzothiadiazole, jasmonic acid (JA), and 1-amino cyclopropane-1-carboxylic acid, which mediate SA-and JA/ethylene (ET)-dependent defense signaling pathways, respectively. Furthermore, the expression of OsBIABP1 is activated by the infection of Magnaporthe oryzae, and the induced expression is quicker and stronger during early stages of pathogenesis in incompatible interaction than that in compatible interaction between rice and M. oryzae. Our results suggest that OsBIABP1 may be a defense-related AMP-binding protein that is involved in the regulation of defense re-sponse through SA and/or JA/ET signaling pathways. 相似文献
12.
对近年来甲壳动物卵黄蛋白原基因的分子克隆和该基因在体内表达研究等方面进行分析,显示甲壳动物卵黄蛋白原的cDNA及其推断的氨基酸序列有很高的相似性,且有类似的分裂位点.卵巢和肝胰腺是甲壳动物卵黄蛋白原基因表达的主要位点. 相似文献
13.
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
14.
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
15.
以cDNA微阵列检测313个已知基因在萌发期水稻幼苗中的表达谱 总被引：1，自引：0，他引：1
使用含512个已知水稻基因3′表达序列标签的cDNA微阵列检测了萌发期水稻幼苗的基因表达谱，313个基因产生了可靠的杂交信号．其中，天冬氨酸氨基转移醇基因和4个具有核糖体功能的基因表达丰度非常高，表明萌发期幼苗中的氨基酸和蛋白质的合成代谢很活跃．β—l，3一葡聚糖酶基因是一种典型的病程相关基因，该基因的高水平表达，表明幼苗中存在着一种高度发育的抗病机制．实验也发现编码一种重要的抑制细胞凋亡的基因-Bax inhibitor-1，在幼苗中的表达丰度很高；该结果可解释为什么正常的幼苗中很少发生细胞程序性死亡现象．实验所测定的大量基因的表达丰度有助于从基因组转录水平理解萌发幼苗的生理特点． 相似文献
16.
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
17.
基因表达分析如定量PCR、RNA印记等需要应用内参基因对目标基因表达量进行校正,以期获得真实可靠的结果,管家基因是常用的内参基因。热带兰金钗石斛中管家基因的研究至今尚无报道。该文对金钗石斛腋芽cDNA文库中的5个管家基因的序列和基因表达进行了分析和检测。其中,4个基因编码泛素蛋白,1个编码a-微管蛋白,分别命名为DnUBQ1、DnUBQ2、DnUBQ3、DnUBQ4及DnTUA1.DnUBQ1基因的密码使用频率表现出与金钗石斛普遍密码子不同的偏好性。在低温春化前后和春化过程中,5个基因的表达均无明显改变,表现为组成型表达。因此,5个基因均可在研究金钗石斛春化相关基因的表达时作为参照基因使用。 相似文献
18.
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
19.