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从菊芋(Helianthus tuberosus L.)块茎cDNA文库中得到了一个新的植物类金属硫蛋白基因htMT2的cDNA序列,全长509 bp,包括240 bp的开放阅读框、62 bp的5′端非翻译区、207 bp的3′端非翻译区.通过PCR获得了2个htMT2编码区的部分基因组片段htMTG-1及htMTG-2,长度分别为986 bp和982 bp.分析表明两个基因组片段均包含3个外显子及2个内含子,编码一个由79个氨基酸残基组成的多肽,与从htMT2推测的多肽完全一致,该多肽具有植物类金属硫蛋白的典型结构特征,N端及C端结构域富含Cys,分别具有8个和7个Cys残基,上述两个结构域被一个无Cys的中间区分开.Southern杂交结果表明,htMT2在菊芋基因组中以小基因家族的形式存在.Northern杂交结果表明htMT2在叶片、叶柄、茎及块茎中均有表达,在茎中有较高水平的表达,但在根中未检测到杂交信号.经Cu2+处理后,htMT2在茎中的表达量显著降低.与其他2型金属硫蛋白的序列同源性比较及htMT2对金属离子处理的反应均表明,htMT2是一种新的植物类金属硫蛋白基因. 相似文献
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Beibei Huang Xiaojun Liu Xinglong Wang Yan Pi Juan Lin Jiong Fei Xiaofen Sun Kexuan Tang 《Molecular Biology》2005,39(5):684-695
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A cDNA encoding a phosphoinositide-specific phospholipase C (PI-PLC) has been isolated from Zea mays by screening a cDNA library. The cDNA, designated ZmPLC, encodes a polypeptide of 586 amino acids, containing the catalytic X, Y and C2 domains found in all PI-PLCs from plants. Northern blot analysis showed that the expression of the ZmPLC gene in roots is up-regulated under conditions of high salt, dehydration, cold or low osmotic stress conditions. Recombinant ZmPLC protein was expressed in Esch- erichia coli, purified and used to produce polyclonal antibody, this polyclonal antibody is important for further studies to assess the ultimate function of the ZmPLC gene in plants. 相似文献
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Xiaojing Xu Yijun Zhou Shanjun Wei Dongtao Ren Min Yang Huahu Bu Mingming Kang Junli Wang Jinchao Feng 《Molecules and cells》2009,27(4):423-428
Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally
occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had
a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein
was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium
cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated
that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic
acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological
pathways activated by a variety of environmental conditions.
These authors contributed equally to this work. 相似文献
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Ladhar-Chaabouni R Mokdad-Gargouri R Denis F Hamza-Chaffai A 《Molecular biology reports》2009,36(5):1007-1014
cDNA probes have been developed for subsequent use in monitoring the cadmium exposure of the clam Ruditapes decussatus and the cockle Cerastoderma glaucum using metallothionein (MT) gene expression in different tissues of these species. Two partial MT cDNAs were isolated from
Ruditapes decussatus and Cerastoderma glaucum. The identification of the nucleotide sequences showed that the cDNAs consist of 480 bp coding 72 amino acid proteins containing
21 cysteine residues organized in Cys–X–Cys motifs as classically described for MTs. The induction of MT gene expression in
CdCl2 treated bivalves was confirmed by dot blot analysis and suggests a potential specific tissue expression rate. 相似文献
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Gholizadeh A Kohnehrouz BB Santha IM Lodha ML Kapoor HC 《Biochemistry. Biokhimii?a》2005,70(9):1005-1010
A small cDNA fragment containing a ribosome-inactivating site was isolated from the leaf cDNA population of Celosia cristata by polymerase chain reaction (PCR). PCR was conducted linearly using a degenerate primer designed from the partially conserved peptide of ribosome-inactivating/antiviral proteins. Sequence analysis showed that it is 150 bp in length. The cDNA fragment was then cloned in a bacterial expression vector and expressed in Escherichia coli as a ~57 kD fused protein, and its presence was further confirmed by Western blot analysis. The recombinant protein was purified by affinity chromatography. The purified product showed strong antiviral activity towards tobacco mosaic virus on host plant leaves, Nicotiana glutinosa, indicating the presence of a putative antiviral determinant in the isolated cDNA product. It is speculated that antiviral site is at, or is separate but very close to, the ribosome-inactivating site. We nominate this short cDNA fragment reported here as a good candidate to investigate further the location of the antiviral determinants. The isolated cDNA sequence was submitted to EMBL databases under accession number of AJ535714. 相似文献
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Fu W Han B Duan D Liu W Wang C 《Journal of industrial microbiology & biotechnology》2008,35(8):915-922
Marine bacterium Vibrio sp. F-6, utilizing agarose as a carbon source to produce agarases, was isolated from seawater samples taken from Qingdao, China. Two agarases (AG-a and AG-b) were purified to a homogeneity from the cultural supernatant of Vibrio sp. F-6 through ammonium sulfate precipitation, Q-Sepharose FF chromatography, and Sephacryl S-100 gel filtration. Molecular weights of agarases were estimated to be 54.0 kDa (AG-a) and 34.5 kDa (AG-b) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH values for AG-a and AG-b were about 7.0 and 9.0, respectively. AG-a was stable in the pH range of 4.0-9.0 and AG-b was stable in the pH range of 4.0-10.0. The optimum temperatures of AG-a and AG-b were 40 and 55 degrees C, respectively. AG-a was stable at temperature below 50 degrees C. AG-b was stable at temperature below 60 degrees C. Zn(2+), Mg(2+) or Ca(2+) increased AG-a activity, while Mn(2+), Cu(2+) or Ca(2+) increased AG-b activity. However, Ag(+), Hg(2+), Fe(3+), EDTA and SDS inhibited AG-a and AG-b activities. The main hydrolysates of agarose by AG-a were neoagarotetraose and neoagarohexaose. The main hydrolysates of agarose by AG-b were neoagarooctaose and neoagarohexaose. When the mixture of AG-a and AG-b were used, agarose was mainly degraded into neoagarobiose. 相似文献
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Summary
Acmella radicans var. radicans propagation was established in vitro. This plant belongs to the Asteraceae from which some species are known for their insecticide, fungicide and antibacterial
activity. The complete Murashige and Skoog (MS) medium was the best in assisting seed germination. In order to obtain shoots
in vitro, a complete MS medium and half-strength MS medium were assayed with explants from leaves, nodes, and internodes. The best
medium for shoot production was the half-strength MS medium with no addition of plant growth regulators, and the highest shoot
propagation was from single-node explants. Regeneration of roots on shoot explants in the medium was obtained without the
addition of growth regulators. Of the plantlets that were acclimated, 90% of them were obtained from rooted shoots with completely
expanded leaves. The alkamide content was evaluated for each tissue and the higher concentration was observed in flower heads.
The main alkamides present in the leaves and the flower heads were N-(2-phenylethyl)-2Z,4E-octadienamide and 3-phenyl-N-(2-phenylethyl)-2-propenamide. This study describes the methodology for the establishment and propagation of Acmella radicans in vitro and the evaluation of different tissue alkamide contents in vitro and in the field. 相似文献
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cDNA cloning and expression analysis of a mannose-binding lectin from <Emphasis Type="Italic">Pinellia pedatisecta</Emphasis> 总被引:2,自引:0,他引:2
Pinellia pedatisecta agglutinin (PPA) is a very basic protein that accumulates in the tuber of P. pedatisecta. PPA is a hetero-tetramer protein of 40 kDa, composed of two polypeptide chains A (about 12 kDa) and two polypeptides chains
B (about 12 kDa). The full-length cDNA of PPA was cloned from P. pedatisecta using SMART RACE-PCR technology; it was 1146 bp and contained a 771 bp open reading frame (ORF) encoding a lectin precursor
of 256 amino acid residues with a 24 amino acid signal peptide. The PPA precursor contained 3 mannose-binding sites (QXDXNXVXY)
and two conserved domains of 43% identity, PPA-DOM1 (polypeptides A) and PPA-DOM2 (polypeptides B). PPA shared varying identities, ranging from 40% to 85%, with mannose-binding lectins from other species
of plant families such as Araceae, Alliaceae, Iridaceae, Liliaceae, Amaryllidaceae and Bromeliaceae. Southern blot analysis
indicated that ppa belonged to a multi-copy gene family. Expression pattern analysis revealed that ppa expressed in most tested tissues, with high expression being found in spadix, spathe and tuber. Cloning of the ppa gene not only provides a basis for further investigation of its structure, expression and regulatory mechanism, but also
enables us to test its potential role in controlling pests and fungal diseases by transferring the gene into plants in the
future. 相似文献