共查询到20条相似文献，搜索用时 15 毫秒
1.
Qin LU Jian-ping LU Xiao-dong LI Xiao-hong LIU Hang MIN Fu-cheng LIN 《Journal of Zhejiang University. Science. B》2008,9(7)
In this study the MTP1 gene, encoding a type Ⅲ integral transmembrane protein, was isolated fi'om the rice blast fungus Magnaporthe oryzae. The Mtpl protein is 520 amino acids long and is comparable to the Ytpl protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP (green fluorescent protein) fusion expression results indicate that Mtpl 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 ex-pressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for patho-genicity. The △mtpl 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
2.
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
3.
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
4.
1IntroductionRibosome-inactivating proteins(RIPs)occur natu-rally in a variety of higher plant species,and theyfunction by catalytic depurination of a specific aden-osine residue located near the3*terminus of eukary-otic large ribosomal subunit rRNA,preventing EF-2/GTP binding and thereby blocking peptidyl-tRNAtranslocation during protein synthesis[1].Many RIPsare potent antiviral and antifungal proteins in vitro[2,3],but it may beinsufficient for field application,as com-pared to the ac… 相似文献
5.
TIAN Yong LU Li-zhi FU Yan TAO Zheng-rong SHEN Jun-da WANG De-qian YUAN Ai-ping YIN Zhao-zheng 《Journal of Zhejiang University. Science. B》2007,8(5)
The protein encoded by CC chemokine receptor 7 (CCR7) is a member of the G protein-coupled receptor family. This receptor was identified as a gene induced by the Epstein-Barr virus (EBV), and is thought to be a mediator of EBV effects on B lymphocytes. This receptor is expressed in various lymphoid tissues and activates B and T lymphocytes. It has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. To map the CCR7 gene in chicken chromosome, a 6 000 rads chicken-hamster radiation hybrid panel (ChickRH6) was used. PCR of samples from ChickRH6 revealed that the location of CCR7 gene is linked to the maker SEQ0347 (6 cR away) with LOD score of 16.6 and that the marker SEQ0347 is located on chromosome 27 at 27 cR of RH (radiation hydrid) map. We compared the corresponding human mRNA sequence with the predicted coding sequence of chicken CCR7 gene, and found that the assembled contig shared a high percentage of similarity with that of the human gene. 相似文献
6.
根据GenBank数据库中猪圆环病毒Ⅱ型的基因组序列设计引物,采用PCR技术从病料基因组DNA中扩增出PCVⅡ河南地方株ORF4基因,全长180 bp,编码59个氨基酸.将该基因克隆至载体pGEX-4T-3中形成pGEX-4T-3-ORF4表达载体.经PCR、酶切和测序鉴定后,转化表达菌株BL21（DE3）诱导表达.SDS-PAGE结果显示：ORF4能够在大肠杆菌中表达,产物的分子量约为32 kD,且以包涵体形式存在.Western Blot检测结果显示,纯化后的ORF4蛋白能够与鼠抗6＆#215;His标签单克隆抗体发生特异性反应,为进一步研究PCVⅡORF4蛋白的特性与功能奠定了基础. 相似文献
7.
根据已发表的序列，通过PCR技术克隆了一系列构建烟草叶绿体多顺反子表达载体所需的元件：质体核搪体（16S）RNA操纵元启动子（Prrn）、质体面A基因3’端（psbA3’）、山菠菜甜菜碱醛脱氢酶基因（BADH）、烟草叶绿体高频同源重组片段（psaA／psbC，大小3463bp）、甘露聚糖酶基因（man）、绿荧光蛋白基因（gfp）。构建了烟草质体多顺反子定点整合表达载体pLM7（-psaA-Prrn-SD-man-SD-gfp-SD-BADH-PSBA3’-PSBC-）。并在大肠杆菌中通过平板定性分析等方法对所构建载体上的表达盒进行了功能鉴定。 相似文献
8.
9.
10.
Assignment of <Emphasis Type="Italic">CCR7</Emphasis> gene to chicken chromosome 27 by radiation hybrid panel mapping 总被引：1，自引：0，他引：1
Tian Y Lu LZ Fu Y Tao ZR Shen JD Wang DQ Yuan AP Yin ZZ 《Journal of Zhejiang University. Science. B》2007,8(5):314-317
The protein encoded by CC chemokine receptor 7 （CCR7） is a member of the G protein-coupled receptor family. This receptor was identified as a gene induced by the Epstein-Barr virus （EBV）, and is thought to be a mediator of EBV effects on B lymphocytes. This receptor is expressed in various lymphoid tissues and activates B and T lymphocytes. It has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. To map the CCR7 gene in chicken chromosome, a 6000 rads chicken-hamster radiation hybrid panel （ChickRH6） was used. PCR of samples from ChickRH6 revealed that the location of CCR7 gene is linked to the maker SEQ0347 （6 cR away） with LOD score of 16.6 and that the marker SEQ0347 is located on chromosome 27 at 27 cR of RH （radiation hydrid） map. We compared the corresponding human mRNA sequence with the predicted coding sequence of chicken CCR7 gene, and found that the assembled contig shared a high percentage of similarity with that of the human gene. 相似文献
11.
INTRODUCTION Soil-borne pathogens, including Pythium spp. and Fusarium spp., cause significant yield losses in horticulture and agriculture crops (Mao et al., 1997). Current practices for controlling plant diseases are based largely on disease resistant crops, cultivation management in fields and application of synthetic pesticides (Elizabeth and Emmert, 1999). Biological control using antagonistic microbes to reduce the use of chemical pesticides in a system of integrated plantdisease … 相似文献
12.
大豆β-1,3-葡聚糖酶基因转化烟草及抗病性的研究 总被引：6，自引：0，他引：6
构建了大豆β－１，３－葡聚糖酶（β－１，３－ｇｌｕｃａｎａｓｅ）基因的植物表达载体ｐＢＩ１２１－ｇｌｕ，并通过直接转化方法将其导入根癌农杆菌（Ａ．ｔｕｍｅｆａｃｉｅｎｓ）ＬＢＡ４４０４受体菌中，构建了用于植物遗传转化的工程菌株ＬＢＡ４４０４（ｐＢＩ１２１－ｇｌｕ），并以烟草为转化对象进行了遗传转儿，获得了大量再生的转基因烟草．ＰＣＲ，ＰＣＲ－Ｓｏｕｔｈｅｍ以及Ｓｏｕｔｈｅｍ杂交检测结果表明目的基因已整合到烟草基因组中．转基因植株苗期抗立枯病实验表明，部分转化β－１，３－葡聚糖酶基因的工程烟草对立枯丝核菌（Ｒｈｉｚｏｃｔｏｎｉａ ｓｏｌａｎｉ．）表明出不同程度的抗性提高． 相似文献
13.
14.
A fiberless seed mutant (fl) was identified in a commercial cotton (Gossypium hirsutum L.) variety Xu-Zhou 142 (FL). This phenotype is associated with lack of fiber cell initiation in the outer integument of
the ovule, as was characterized by analysis of genes related to fiber differentiation and development. Two genes, fl-E6 and
FL-E6, were cloned from fl-integument cells and FL-fiber or integument cells, respectively. Compared with FL-E6, fl-E6 showed
a dramatic change in nucleotide sequence: (1) FL-E6 contained a tandem repetitive sequence in which GGCTCA (Gly-Ser) is repeated
five times between the 82nd and the 93rd codon from the first ATG codon, while in fl-E6 the same sequence is repeated four
times; (2) The fl-E6 gene encodes a polypeptide of 241 amino acids but lacks two codons between the 90th and 93rd codon and
three between the 171st and 174th relative to FL-E6; (3) There are also 12 nucleotide substitutions which would result in
7 amino acid differences between fl-E6 and FL-E6. Analysis of RT-PCR and Northerm Blot showed that expression of the fl-E6
gene is suppressed in the fl-integument cells, but highly expressed in FL-fiber cells. The difference between fl-E6 and FL-E6
may be associated with lower expression of fl-E6 in the fl-integument cells. Searches of protein databases with the FL-E6
gene sequence showed similarity to the protein backbones of two arabinogalactan-proteins (AGPs), one from the filtrate of
suspension-cultured cells ofPyrus cornmunis (AGPPc2) and the other fromNicotiana alata (AGPNa2). Although the function of the FL-E6 protein in differentiation and development of cotton fiber cells is not known,
the data indicate that the mutation of fl-E6 gene from FL-E6 gene may inhibit the fiber cell initiation from epidermal cells
of the outer integument of the ovule.
Project supported by the National Natural Science Foundation of China (Nos. 39770473 and 30170058), National Program of Plant
Gene Transfer (J00-B-002-10), National High Technology Research and Development (863) Program of China (No. 2001AA212081),
and Research Program of Ministry of Education of China (No. 0114) 相似文献
15.
[目的]新的大肠癌相关性抗原EID3的基因克隆及其诊断价值研究.[方法]利用大肠癌病人体内血清中所含的对肿瘤抗原产生的特异性抗体筛选睾丸组织cDNA噬菌体表达文库和大肠癌组织cDNA噬菌体表达文库(SEREX),并用RT-PCR技术研究EID3 mRNA在正常组织和大肠癌传代细胞表达.[结果]睾丸组织cDNA噬菌体表达文库筛选得到了可以诱导大肠癌病人抗体免疫应答的新抗原EID3基因(Gen-bank NM_001008394.1).它们定位于染色体19q13.2,EID3含1个外显子.通过RT-PCR分析发现,EID3基因在43例大肠癌传代细胞株中,39例阳性,阳性率为90.7%.在正常组织中,除睾丸组织外不表达或有极低水平转录.[结论]EID3 mRNA表达检测用于诊断大肠癌,可能具有高特异性和高敏感性的特点.EID3蛋白被首次发现在大肠癌病人中能够诱导机体的抗体免疫应答,为一个新的大肠癌相关性抗原分子.其功能可能与抑制细胞的恶性增殖相关,并可进一步研究其用于治疗和诊断大肠癌的可行性. 相似文献
16.
Zhang Xin Zhang Bing-xin Zhang Zhen Shen Wei-feng Yang Ching-hong Yu Jing-quan Zhao Yu-hua 《Journal of Zhejiang University. Science. B》2005,6(8):770-777
Fusarium head blight (FHB) caused byFusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing
plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents,Brevibacillus brevis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of theBrevibacillus brevis ZJY-1 andBacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field
study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved
GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both
biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley
FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation withF. graminearum.
Project supported by the National Natural Science Foundation of China (No. 30230250) and Science and Technology Committee
of Zhejiang Province (No. 2003C22029), China 相似文献
17.
研究用亲和融合谷胱甘肽 S 转移酶 (GST)的方法纯化重组人白细胞介素 6(IL 6)的发酵和纯化工艺 ,使用含有质粒pHZl818的E .coliJMl0 9在 2XYT培养基中进行发酵表达 ,IL 6表达为与谷胱甘肽 S 转移酶 (GST)融合的融合蛋白GST IL 6.融合蛋白存在可溶的活性蛋白和不可溶的包含体两种形式 ,此包含体无活性且无法复性 ,无法用亲和层析回收 ,通过实验优化摇床发酵的诱导温度和转速 ,以增加可溶融合蛋白的表达 .菌体超声破碎液后 ,上清液用作亲和柱层析 ,可将融合蛋白提纯至 80 00 ,每升发酵液可得 10mg融合蛋白 ,用凝血酶裂解处理 6h ,亲和标志物GST被特异性切除 ,裂解得到的IL 6用离子交换柱层析可纯化至 95 00 ,MTT法测得纯化的IL 6生物学活性为 1.0 2× 10 8IU/mg . 相似文献
18.
从假单胞菌（Pseudomonassp．）XZG36中克隆弹性蛋白酶基因,构建原核表达载体,实现其在大肠杆菌（Escherichiacoli）中的高效表达,并对表达产物进行酶学性质分析,为微生物发酵生产弹性蛋白酶奠定基础．以假单胞菌基因组DNA为模板,PCR扩增弹性蛋白酶基因,并将其开放阅读框（0RF）克隆至融合表达载体pET30a（＋）进一步IPTG诱导表达;表达产物经His·Bind亲和层析纯化后对弹性蛋白酶进行酶学性质分析．实验成功克隆了弹性蛋白酶基因,DNA基因片段为1672bp、编码497个氨基酸残基的多肽,与预计长度相符合;实现了其在E．coli中的高效表达,表达量约占菌体总蛋白的20％;经SDS-PAGE分析,相对分子质量为48000,与预期的一致;提纯后的表达蛋白SDS-PAGE分析可见单一条带,纯度可达92％以上．表达蛋白具有良好活性． 相似文献
19.
余瑛 《重庆大学学报(英文版)》2003,2(2)
1. Introduction Acidic fibroblast growth factor (aFGF), a member of a family of structurally related polypeptides, is a kind of multifunctional protein that can stimulate cellular proliferation, migration and differentiation. In vivo, aFGF displays angiogenic activity promoting vascular endothelial cell mitogenesis as well as chemotaxis and induction of proteases involved in tissue regeneration [1]. Due to this wide range of biological activities, many potential therapeutic usages of aFGF … 相似文献
20.
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