共查询到20条相似文献,搜索用时 31 毫秒
1.
Xiangbin Xu Jufang Bian Songbai Liu Hongmiao Song Nongnong Shi Yuezhi Tao Huizhong Wang 《Molecular breeding : new strategies in plant improvement》2011,27(3):337-346
The PROMOTION OF CELL SURVIVAL 1 (PCS1) gene, encoding an aspartic protease, has an important role in determining the fate of cells in embryonic development and
reproduction processes in Arabidopsis. To explore the potential function of the PCS1 gene in generating reproductive sterility, we placed the PCS1 gene under the control of an 1,869-bp nucleotide sequence from the 3′ end of the second intron (AG-I) of Arabidopsis AGAMOUS and CaMV 35S (–60) minimal promoter [AG-I-35S (–60)::PCS1], and introduced it into tobacco. RT–PCR results demonstrated that the PCS1 gene driven by AG-I-35S (–60) chimeric promoter was expressed only in anthers and carpels in the reproductive tissues of transgenic tobacco. Compared to
wild-type plants, all AG-I-35S (–60) and AG-I-35S (–60)::PCS1 transgenic lines showed a normal phenotype throughout the vegetative growth phase. However, during the reproductive stage,
most AG-I-35S (–60)::PCS1 transgenic plant anthers displayed delayed dehiscence, failed dehiscence, petalody and hypoplasia, and the pollen grains
had different shapes and sizes with a distorted, shrunken, or collapsed morphology. Moreover, three transgenic lines, PCS1-1,
PCS1-3 and PCS1-4, showed higher sterility than wild-type and AG-I-35S (–60) transgenic plants, respectively. These results showed that the construct of AG-I-35S (–60)::PCS1 was partially effective at preventing seed set and provided a novel sterility strategy. 相似文献
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A major limitation on the expression of some foreign proteins in transgenic plants is the toxic effect of such proteins on
the host plant resulting in inhibition of normal growth and development. A solution to this problem is to control the expression
of genes for such proteins by means of inducible promoters, as is frequently done in microbial systems. A cDNA clone was obtained
from subtractive hybridization of non-harvested and harvested alfalfa leaf tissue, named hi12. The hi12 cDNA was identified as part of the S-adenosyl-l-methionine: trans-caffeoyl-CoA3-O-methyltransferase gene of alfalfa, a gene encoding an essential key enzyme in lignin synthesis. The hi12 gene was strongly induced by harvesting and wounding but not by heat shock. The promoter of the hi12 gene, isolated by genomic walking, contained several stress response cis-elements. Transgenic plants of tobacco and Medicago truncatula containing the GUS gene driven by the promoter showed GUS expression following harvesting, demonstrating the activity of
these regulatory regions in other plant species. 相似文献
4.
Kimyung Choi Joohyun Shim Nayoung Ko Heejong Eom Jiho Kim Jeong-Woong Lee Dong-Il Jin Hyunil Kim 《Transgenic research》2017,26(2):209-224
Production of transgenic pigs for use as xenotransplant donors is a solution to the severe shortage of human organs for transplantation. The first barrier to successful xenotransplantation is hyperacute rejection, a rapid, massive humoral immune response directed against the pig carbohydrate GGTA1 epitope. Platelet activation, adherence, and clumping, all major features of thrombotic microangiopathy, are inevitable results of immune-mediated transplant rejection. Human CD39 rapidly hydrolyzes ATP and ADP to AMP; AMP is hydrolyzed by ecto-5′-nucleotidase (CD73) to adenosine, an anti-thrombotic and cardiovascular protective mediator. In this study, we developed a vector-based strategy for ablation of GGTA1 function and concurrent expression of human CD39 (hCD39). An hCD39 expression cassette was constructed to target exon 4 of GGTA1. We established heterozygous GGTA1 knock-out cell lines expressing hCD39 from pig ear fibroblasts for somatic cell nuclear transfer (SCNT). We also described production of heterozygous GGTA1 knock-out piglets expressing hCD39 and analyzed expression and function of the transgene. Human CD39 was expressed in heart, kidney and aorta. Human CD39 knock-in heterozygous ear fibroblast from transgenic cloned pigs, but not in non-transgenic pig’s cells. Expression of GGTA1 gene was lower in the knock-in heterozygous ear fibroblast from transgenic pigs compared to the non-transgenic pig’s cell. The peripheral blood mononuclear cells (PBMC) from the transgenic pigs were more resistant to lysis by pooled complement-preserved normal human serum than that from wild type (WT) pig. Accordingly, GGTA1 mutated piglets expressing hCD39 will provide a new organ source for xenotransplantation research. 相似文献
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Yunyun Cheng Songcai Liu Xin Zhang Qingyan Wu Siming Li Haoyu Fu Lijie Dong Hao Yu Linlin Hao 《The protein journal》2016,35(3):231-236
Insulin like growth factor 1 receptor (IGF-1R) is a candidate gene for growth and carcass traits in regulating animal growth, metabolism and endocrine. It is widely expressed in liver, muscle, bone tissues where the IGF-1R functions as a factor that promotes cell growth. In this study, the protein expression level of IGF-1R gene in liver and muscle tissues of three periods (birth, weaning and adult) of three pig breeds (BamaXiang pigs (BM), Tibetan pigs (TM) and Junmu No.1 pigs (JM)) were tested by western blot. SNPs within the regulatory region of pig IGF-1R gene were detected using direct sequencing and then the genotypes were identified through AS-PCR approach. Results showed expression profiles of IGF-1R gene between liver and muscle tissues were different and significant differences were also found among pig breeds. In the same time, four SNPs were detected in the regulatory region of IGF-1R gene, among which the genotype frequency of three (g.?1468G > C, g.?1192 C > T and g.330,424 C > T) were significantly different among the pig breeds. BM tended to heterozygous (GC/CT) of the anterior two loci, while TM and JM preferred the other two homozygotes respectively. For the g.330,424 C > T, all pig breeds were tended to be the heterozygous. In conclusion, the SNPs with different genotype distribution among the three pig breeds may explain the gene expression difference between the different pig breeds. 相似文献
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Modulatory Effects of <Emphasis Type="Italic">Lactobacillus salivarius</Emphasis> on Intestinal Mucosal Immunity of Piglets 总被引:2,自引:0,他引:2
Recent studies have demonstrated that lactobacilli or their cell components can improve certain immune function in animals.
The aim of this study is to investigate the effects of porcine lactobacilli on the intestinal mucosal immunity of piglets.
Neonatal piglets were used as a model and were orally administrated with Lactobacillus salivarius B1 isolated from the duodenal mucosa of a healthy piglet. The feces of the piglets were collected on days 7, 14, and 21 for
intestinal microflora analysis. On day 28, the piglets were sacrificed, and their intestinal mucosa samples were immediately
collected to investigate the changes in intestinal morphological and immunocompetent cells. Finally, the expression of cytokines
and TLRs was detected in the different intestinal segments. The results indicate that L. salivarius B1 can partially ameliorate the microflora of the feces and increase the number of intestinal immunocompetent cells, as the
intraepithelial lymphocyte (P < 0.05), and the IgA-producing cells (P < 0.01) in the lactobacilli-treated group were all increased compared with those in the control group. Enhanced expression
of the cytokine IL-6 gene was also observed in the ileum (P < 0.05). Moreover, L. salivarius B1 can also upregulate the expression of TLR2 in the intestinal tract at the gene and protein levels (P < 0.05). The results demonstrate that L. salivarius B1 is beneficial for the maturation of the intestinal mucosal immune system and elicited local immunomodulatory activities.
In addition, the modulatory effects of L. salivarius B1 on mucosal immunity mainly depend on its extracellular components. 相似文献
8.
Mohsin Abbas Zaidi Gongyin Ye Hongwei Yao Taek H. You Evelin Loit Donald H. Dean Sheikh Riazuddin Illimar Altosaar 《Molecular biotechnology》2009,43(3):232-242
Nucleotide sequence encoding the truncated insecticidal Cry1Ca1 protein from Bacillus thuringiensis was extensively modified based on the codon usage of rice genes. The overall G + C contents of the synthetic cry1Ca1 coding sequence were raised to 65% with an additional bias of enriching for G and C ending codons as preferred by monocots.
The synthetic gene was introduced into the Chinese japonica variety, Xiushui 11, by Agrobacterium-mediated transformation. Transgenic rice plants harboring this gene were highly resistant to Chilo
suppressalis and Spodoptera litura larvae as revealed by insect bioassays. High levels of Cry1Ca1 protein were obtained in the leaves of transgenic rice, which
were effective in achieving 100% mortality of S. litura and C. suppressalis larvae. The levels of Cry1Ca1 expression in the leaves of these transgenic plants were up to 0.34% of the total soluble proteins.
The larvae of C. suppressalis and S.
litura could consume a maximum of 1.89 and 4.89 mm2 of transgenic leaf area whereas the consumption of non-transgenic leaves by these larvae was significantly higher; 58.33 and
61.22 mm2, respectively. Analysis of R1 transgenic plants indicated that the cry1Ca1 was inherited by the progeny plants and provided complete protection against C. suppressalis and S.
litura larvae. 相似文献
9.
An SJ Pandeya D Park SW Li J Kwon JK Koeda S Hosokawa M Paek NC Choi D Kang BC 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,122(3):459-470
A temperature-sensitive mutant of Capsicum chinense, sy-2, shows a normal developmental phenotype when grown above 24°C. However, when grown at 20°C, sy-2 exhibits developmental defects, such as chlorophyll deficiency and shrunken leaves. To understand the underlying mechanism
of this temperature-dependent response, phenotypic characterization and genetic analysis were performed. The results revealed
abnormal chloroplast structures and cell collapse in leaves of the sy-2 plants grown at 20°C. Moreover, an excessive accumulation of reactive oxygen species (ROS) resulting in cell death was detected
in the chlorophyll-deficient sectors of the leaves. However, the expression profile of the ROS scavenging genes did not alter
in sy-2 plants grown at 20°C. A further analysis of fatty acid content in the leaves showed the impaired pathway of linoleic acid
(18:2) to linolenic acid (18:3). Additionally, the Cafad7 gene was downregulated in sy-2 plants. This change may lead to dramatic physiological disorder and alteration of leaf morphology in sy-2 plants by losing low-temperature tolerance. Genetic analysis of an F2 population from a cross between C. chinense ‘sy-2’ and wild-type C. chinense ‘No. 3341’ showed that the sy-2 phenotype is controlled by a single recessive gene. Molecular mapping revealed that the sy-2 gene is located at a genomic region of the pepper linkage group 1, corresponding to the 300 kb region of the Ch1_scaffold
00106 in tomato chromosome 1. Candidate genes in this region will reveal the identity of sy-2 and the underlying mechanism of the temperature-dependent plant response. 相似文献
10.
Raham Sher Khan Syed Sartaj Alam Iqbal Munir Pejman Azadi Ikuo Nakamura Masahiro Mii 《Plant Cell, Tissue and Organ Culture》2011,106(1):11-20
The presence of marker genes conferring antibiotic or herbicide resistance in transgenic plants has been a controversial issue
and a serious problem for their public acceptance and commercialization. The MAT (multi-auto-transformation) vector system
has been one of the strategies developed to excise the selection marker gene and produce marker-free transgenic plants. In
an attempt to produce transgenic marker-free Petunia hybrida plants resistant to Botrytis cinerea (gray mold), we used the ipt gene as a selectable marker gene and the wasabi defensin (WD) gene, isolated from Wasabia japonica (a Japanese horseradish which has been a potential source of antimicrobial proteins), as a gene of interest. The WD gene was cloned from the binary vector, pEKH-WD, to an ipt-type MAT vector, pMAT21, by gateway cloning technology and transferred to Agrobacterium
tumefaciens strain EHA105. Infected leaf explants of P.
hybrida were cultured on hormone- and antibiotic-free MS medium. Extreme shooty phenotype (ESP)/ipt shoots were produced by the explants infected with the pMAT21-WD. The same antibiotic- and hormone-free MS medium was used in subcultures of the ipt shoots. Ipt shoots subsequently produced morphologically normal shoots. Molecular analyses of genomic DNA from the transgenic plants
confirmed the integration of the gene of interest and excision of the selection marker. Expression of the WD gene was confirmed by northern blot and western blot analyses. A disease resistance assay of the marker-free transgenic plants
exhibited enhanced resistance against B. cinerea strain 40 isolated from P. hybrida. 相似文献
11.
Olga A. Shulga Tatyana Yu Mitiouchkina Anna V. Shchennikova Konstantin G. Skryabin Sergey V. Dolgov 《In vitro cellular & developmental biology. Plant》2011,47(5):553-560
Chrysanthemum is one of the most important commercial cut flowers in the world. Early-flowering cultivars are required to
produce quality chrysanthemum flowers with a lower cost of production. To shorten the vegetative growth phase of chrysanthemum,
three AP1-like genes from Asteraceae were constitutively overexpressed in 80 independent transgenic chrysanthemum lines. All lines were characterized by PCR and
RT-PCR and demonstrated that overexpression of compositae AP1-homologs in transgenic chrysanthemum under long-day conditions had no effect on plant development compared to non-transgenic
controls. Conversely, under short-day conditions, transgenic plants commenced bud initiation 2 wk earlier than non-transgenic
chrysanthemum plants. Subsequently, transgenic chrysanthemum flowers showed color earlier and resulted in full opening of
inflorescences 3 wk prior to non-transgenic control plants. These results open new possibilities for genetic improvement and
breeding of chrysanthemum cultivars. 相似文献
12.
Schetelig MF Nirmala X Handler AM 《Apoptosis : an international journal on programmed cell death》2011,16(8):759-768
Pro-apoptotic proteins from the reaper, hid, grim (RHG) family are primary regulators of programmed cell death in Drosophila due to their antagonistic effect on inhibitor of apoptosis (IAP) proteins, thereby releasing IAP-inhibition of caspases that
effect apoptosis. Using a degenerate PCR approach to conserved domains from the 12 Drosophila species, we have identified the first reaper and hid orthologs from a tephritid, the Caribfly Anastrepha
suspensa. As-hid is the first identified non-drosophilid homolog of hid, and As-rpr is the second non-drosophilid rpr homolog. Both genes share more than 50% amino acid sequence identity with their Drosophila homologs, suggesting that insect pro-apoptotic peptides may be more conserved than previously anticipated. Importantly, both
genes encode the conserved IBM and GH3 motifs that are key for IAP-inhibition and mitochondrial localization. Functional verification
of both genes as cell death effectors was demonstrated by cell death assays in A. suspensa embryonic cell culture, as well as in heterologous Drosophila melanogaster S2 cells. Notably, heterologous cell death activity was found to be higher for Anastrepha genes than their Drosophila counterparts. In common with the Drosophila cognates, As-hid and As-rpr negatively regulated the Drosophila inhibitor of apoptosis (DIAP1) gene to promote apoptosis, and both genes when used together effected increased cell death
activity, indicating a co-operative function for As-hid and As-rpr. We show that these tephritid cell death genes are functional and potent as cell death effectors, and could be used to design
improved transgenic lethality systems for insect population control. 相似文献
13.
Mi-Suk Seo Sakiko Takahashi Koh-ichi Kadowaki Makoto Kawamukai Manabu Takahara Tadashi Takamizo 《Plant Cell, Tissue and Organ Culture》2011,107(2):325-332
Panicum meyerianum Nees is a wild relative of Panicum maximum Jacq. (guinea grass), which is an important warm-season forage grass and biomass crop. We investigated the conditions that
maximized the transformation efficiency of P. meyerianum by Agrobacterium infection by monitoring the expression of the β-glucuronidase (GUS) gene. The highest activities of GUS in calli were achieved
by the co-cultivation of plants with Agrobacterium at 28°C for 6 days. We transferred the ddsA gene, which encodes decaprenyl diphosphate synthase and is required for coenzyme Q10 (CoQ10) synthesis, into P. meyerianum by using our optimized co-cultivation procedure for transformation. We confirmed by PCR and DNA gel blot hybridization that
all hygromycin-resistant plants retained stable insertion of the hpt and ddsA genes. We also demonstrated strong expression of S14:DdsA protein in the leaves of transgenic P. meyerianum. Furthermore, we showed that transgenic P. meyerianum produced CoQ10 at levels 11–20 times higher than that of non-transformants. By comparison, the CoQ9 level in transgenic plants
was dramatically reduced. This is the first report of efficient Agrobacterium-mediated transfer of a foreign gene into the warm-season grass P. meyerianum. 相似文献
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Plants have evolved several defense mechanisms, including resistance genes. Resistance to the root-knot nematode Meloidogyne
incognita has been found in wild plant species. The molecular basis for this resistance has been best studied in the wild tomato Solanum peruvianum and it is based on a single dominant gene, Mi-1.2, which is found in a cluster of seven genes. This nematode attacks fiercely several crops, including potatoes. The genomic
arrangement, number of copies, function and evolution of Mi-1 homologs in potatoes remain unknown. In this study, we analyzed partial genome sequences of the cultivated potato species
S. tuberosum and S. phureja and identified 59 Mi-1 homologs. Mi-1 homologs in S. tuberosum seem to be arranged in clusters and located on chromosome 6 of the potato genome. Previous studies have suggested that Mi-1 genes in tomato evolved rapidly by frequent sequence exchanges among gene copies within the same cluster, losing orthologous
relationships. In contrast, Mi-1 homologs from cultivated potato species (S. tuberosum and S. phureja) seem to have evolved by a birth-and-death process, in which genes evolve mostly by mutations and interallelic recombinations
in addition to sequence exchanges. 相似文献
16.
Buriev ZT Saha S Shermatov SE Jenkins JN Abdukarimov A Stelly DM Abdurakhmonov IY 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2011,123(8):1359-1373
The Gossypium MIC-3 (Meloidogyne Induced Cotton-3) gene family is of great interest for molecular evolutionary studies because of its uniqueness to Gossypium species, multi-gene content, clustered localization, and root-knot nematode resistance-associated features. Molecular evolution
of the MIC-3 gene family was studied in 15 tetraploid and diploid Gossypium genotypes that collectively represent seven phylogenetically distinct genomes. Synonymous (dS) and non-synonymous (dN) nucleotide substitution rates suggest that the second of the two exons of the MIC-3 genes has been under strong positive selection pressure, while the first exon has been under strong purifying selection to
preserve function. Based on nucleotide substitution rates, we conclude that MIC-3 genes are evolving by a birth-and-death process and that a ‘gene amplification’ mechanism has helped to retain all duplicate
copies, which best fits with the “bait and switch” model of R-gene evolution. The data indicate MIC-3 gene duplication events occurred at various rates, once per 1 million years (MY) in the allotetraploids, once per ~2 MY in
the A/F genome clade, and once per ~8 MY in the D-genome clade. Variations in the MIC-3 gene family seem to reflect evolutionary selection for increased functional stability, while also expanding the capacity
to develop novel “switch” pockets for responding to diverse pests and pathogens. Such evolutionary roles are congruent with
the hypothesis that members of this unique resistance gene family provide fitness advantages in Gossypium. 相似文献
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Peng Gao Xi Bai Liang Yang Dekang Lv Xin Pan Yong Li Hua Cai Wei Ji Qin Chen Yanming Zhu 《Molecular biology reports》2011,38(1):237-242
Salinity and alkalinity are the two main environmental factors that limit rice production. Better understanding of the mechanisms
responsible for salinity and alkaline stress tolerance would allow researchers to modify rice to increase its resistance to
salinity and alkaline stress. MicroRNAs (miRNAs) are ~21-nucleotide RNAs that are ubiquitous regulators of gene expression
in eukaryotic organisms. Some miRNAs acts as an important endogenous regulator in plant responses to abiotic stressors. miR393
is a conservative miRNA family that occurs in a variety of different plants. The two members of the miR393 family found in
rice are named osa-MIR393 and osa-MIR393b. We found that the osa-MIR393 expression level changed under salinity and alkaline stress, whereas that of osa-MIR393b did not. Target genes of osa-MIR393 were predicted, and some of these putative targets are abiotic related genes. Furthermore, we generated transgenic rice and
Arabidopsis thaliana that over-expressed osa-MIR393, and the phenotype analysis showed that these transgenic plants were more sensitive to salt and alkali treatment compared
to wild-type plants. These results illustrate that over-expression of osa-MIR393 can negatively regulate rice salt-alkali stress tolerance. 相似文献
19.
Guowen Duan Robert B. Saint Chris A. Helliwell Carolyn A. Behm Peter M. Waterhouse Karl H. J. Gordon 《Transgenic research》2010,19(6):1121-1128
Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We created transgenic strains of Drosophila melanogaster in which the rrf-1 or ego-1 RdRp genes from C. elegans were placed under the control of the yeast GAL4 upstream activation sequence. Activation of the gene was performed by crossing
these lines to flies carrying the GAL4 transgene under the control of various Drosophila enhancers. RT–PCR confirmed the successful expression of each RdRp gene. The resulting phenotypes indicated that introduction
of the RdRp genes had no effect on D. melanogaster morphological development. 相似文献