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61.
62.
Methylation protects miRNAs and siRNAs from a 3'-end uridylation activity in Arabidopsis 总被引:5,自引:0,他引:5
Small RNAs of 21-25 nucleotides (nt), including small interfering RNAs (siRNAs) and microRNAs (miRNAs), act as guide RNAs to silence target-gene expression in a sequence-specific manner. In addition to a Dicer homolog, DCL1, the biogenesis of miRNAs in Arabidopsis requires another protein, HEN1. miRNAs are reduced in abundance and increased in size in hen1 mutants. We found that HEN1 is a miRNA methyltransferase that adds a methyl group to the 3'-most nucleotide of miRNAs, but the role of miRNA methylation was unknown. Here, we show that siRNAs from sense transgenes, hairpin transgenes, and transposons or repeat sequences, as well as a new class of siRNAs known as trans-acting siRNAs, are also methylated in vivo by HEN1. In addition, we show that the size increase of small RNAs in the hen1-1 mutant is due to the addition of one to five U residues to the 3' ends of the small RNAs. Therefore, a novel uridylation activity targets the 3' ends of unmethylated miRNAs and siRNAs in hen1 mutants. We conclude that 3'-end methylation is a common step in miRNA and siRNA metabolism and likely protects the 3' ends of the small RNAs from the uridylation activity. 相似文献
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64.
Bing Xu Shaolin Wang Yan Jiang Linlin Yang Ping Li Caixia Xie 《Animal biotechnology》2013,24(4):217-225
Grass carp, Ctenopharyngodon idellus (Valenciennes, 1844), is an economically important species widely cultured in the world, but its genome research resources are largely lacking. The objectives of this study were to construct normalized cDNA libraries for efficient EST analysis, to generate ESTs from these libraries, and to identify EST-related molecular markers such as microsatellites and single nucleotide polymorphisms (SNPs) for genetic analysis of this species. A total of 6,269 ESTs were generated representing 4,815 unique sequences, from which 105 putative microsatellites and 5,228 SNPs were identified. These genome resources provide the material basis for future genetic and functional analyses in this species. 相似文献
65.
cⅠ857基因的体外定位同义突变陈南春,高辉,陈苏民,杨萍,刘新平(西安第四军医大学分子生物学研究所,西安710032)外源基因要在大肠杆菌中获得高表达,需要合适的SD序列和可调控的强启动子[1]。PL启动子在原核启动子中属强启动子,它受cⅠ基因产物... 相似文献
66.
A simple and sensitive assay for 1-aminocyclopropane-1-carboxylic acid 总被引:99,自引:0,他引:99
A simple, rapid, and sensitive method for the quantitative determination of 1-amino-cyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene in plant tissues, is described. The assay is based on the liberation of ethylene from ACC with NaOCl in the presence of Hg2+; ethylene is assayed by gas chromatography. The yield is normally 80% and can be determined by internal standards. The method is quite specific and can detect as little as 5 pmol of ACC. 相似文献
67.
Cryptochrome blue light photoreceptors share sequence similarity to photolyases, flavoproteins that mediate light-dependent DNA repair. However, cryptochromes lack photolyase activity and are characterized by distinguishing C-terminal domains. Here we show that the signaling mechanism of Arabidopsis cryptochrome is mediated through the C terminus. On fusion with beta-glucuronidase (GUS), both the Arabidopsis CRY1 C-terminal domain (CCT1) and the CRY2 C-terminal domain (CCT2) mediate a constitutive light response. This constitutive photomorphogenic (COP) phenotype was not observed for mutants of cct1 corresponding to previously described cry1 alleles. We propose that the C-terminal domain of Arabidopsis cryptochrome is maintained in an inactive state in the dark. Irradiation with blue light relieves this repression, presumably through an intra- or intermolecular redox reaction mediated through the flavin bound to the N-terminal photolyase-like domain. 相似文献
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An efficient tissue culture system for high frequency of plant regeneration from hypocotyl explants of Brassica carinata was developed via manipulation of culture medium and selection of explants. Explants grown on medium containing combinations of 2 mg l-1 BA and 0.01 mg l-1 NAA or 4 mg l-1 kinetin and 0.01 mg l-1 2,4-D regenerated shoots at 100% frequency. High frequency shoot regeneration occurred only from explants originating from 6 to 7-day-old but not younger or older seedlings. Explants showed higher regeneration capacity at the distal end than the proximal end, and the upper segment was more regenerative than the lower segment of hypocotyl. Regenerants were rooted on half-strength growth regulator-free medium, acclimatized and developed into normal, fertile plants.Abbreviations BA
benzyladenine
- 2-4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- MS
Murashige & Skoog 相似文献
70.
玉米转录因子zmCBF1的凝胶阻滞分析 总被引:2,自引:0,他引:2
凝胶阻滞试验是分析核酸与蛋白质相互作用的有效方法,在转录因子的功能分析中得到了广泛应用。以玉米转录因子zmCBF1与顺式元件CRT的结合为例,建立了用于转录因子分析的GRA/EMSA实验体系,并对其在应用中可能出现的问题进行了分析。 相似文献