树鼩IL-2Rα(CD25)基因的分离、鉴定及表达分析

3%～66.4%.RT-PCR检测发现,tsCD25 mRNA分布于树鼩的外周血、脾脏和肺,并受PMA 和 ionomycin 刺激的调节.我们的结果为下一步制备tsCD25单克隆抗体,抑制CD4+CD25+Tregs功能以及开展有关的研究工作奠定了基础.

Characterization of TRPC2, an Essential Genetic Component of VNS Chemoreception, Provides Insights into the Evolution of Pheromonal Olfaction in Secondary-Adapted Marine Mammals

Pheromones are chemical cues released and sensed by individuals of the same species, which are of major importance in regulating reproductive and social behaviors of mammals. Generally, they are detected by the vomeronasal system (VNS). Here, we first investigated and compared an essential genetic component of vomeronasal chemoreception, that is, TRPC2 gene, of four marine mammals varying the degree of aquatic specialization and related terrestrial species in order to provide insights into the evolution of pheromonal olfaction in the mammalian transition from land to water. Our results based on sequence characterizations and evolutionary analyses, for the first time, show the evidence for the ancestral impairment of vomeronasal pheromone signal transduction pathway in fully aquatic cetaceans, supporting a reduced or absent dependence on olfaction as a result of the complete adaptation to the marine habitat, whereas the amphibious California sea lion was found to have a putatively functional TRPC2 gene, which is still under strong selective pressures, reflecting the reliance of terrestrial environment on chemical recognition among the semiadapted marine mammals. Interestingly, our study found that, unlike that of the California sea lion, TRPC2 genes of the harbor seal and the river otter, both of which are also semiaquatic, are pseudogenes. Our data suggest that other unknown selective pressures or sensory modalities might have promoted the independent absence of a functional VNS in these two species. In this respect, the evolution of pheromonal olfaction in marine mammals appears to be more complex and confusing than has been previously thought. Our study makes a useful contribution to the current understanding of the evolution of pheromone perception of mammals in response to selective pressures from an aquatic environment.

鲢(Hypophthalmichthys molitrix)IL-10基因的克隆及表达分析

白细胞介素10(IL-10)是一种作用广泛的抗炎细胞因子,主要功能是限制和最终终止炎症反应．用RACE(rapid amplification of cDNA ends)-PCR方法扩增出鲢白细胞介素10(IL-10)的 cDNA,其全长为1248nt,包含5’非编码区156 nt,3’非编码区552nt,开放阅读框540nt．鲢IL- 10的开放阅读框编码179个氨基酸,其中包含构成两对二硫键的4个保守半胱氨酸．RT-PCR结果显示鲢IL-10 mRNA主要在脾脏、鳃、头肾和肌肉中表达．将鲢IL-10完

鳜IL-1β基因的克隆及原核表达

用RACE PCR获得了鳜白介素-1β(IL-1β)的全长cDNA.鳜IL-1β的cDNA全长为1298 nt(核苷酸),其5′非编码区包含UTR 93 nt;3′非编码区包含452 nt;其开放阅读框内包含753 nt,翻译成251个氨基酸.将鳜IL-1β克隆到原核表达载体pET-32a上,在大肠杆菌Rosetta- gami(DE3)内以包涵体形式得以高效表达.

草鱼、中华鳖脾细胞培养上清液IL-2物质的检测

用刀豆蛋白A(ConA)刺激诱导草鱼和中华鳖脾细胞 ,收集细胞培养上清液 ,用小鼠胸腺细胞增殖试验和对小鼠L92 9细胞系杀伤试验检测上清液中白细胞介素 2 (简称IL 2 )活性。结果表明 :草鱼、中华鳖脾细胞培养上清液中有IL 2样活性物质 ,这种物质使小鼠的胸腺细胞3H TdR掺入量明显增加 ,在相同效靶比的条件下对小鼠L92 9细胞系杀伤率也显著增强 ,这种IL 2活性均能被抗人rIL 2血清所抑制。中华鳖脾细胞培养上清液 (含IL 2 )对中华鳖胸腺细胞也有较明显的促增殖作用并能消除兔抗中华鳖胸

草鱼、中华鳖IL-2活性影响因素的研究

结果显示 :当年草鱼种和中华鳖脾细胞培养上清液中能检测出IL 2活性 ,而且 1龄以上草鱼、中华鳖的IL 2活性高于当年孵化的草鱼和中华鳖。草鱼、中华鳖脾细胞在 2 5℃培养温度条件下 ,其上清液中IL 2活性最高 ,35℃次之 ,1 5℃最低。因此 ,草鱼、中华鳖IL 2活性在一定范围内是随着年龄增加而增强和依赖温度的。通过小鼠胸腺细胞增殖和对小鼠L92 9细胞杀伤率的实验表明 37℃比 2 5℃检测温度下所测的IL 2活性高 ,而中华鳖胸腺细胞增殖实验却显示 2 5℃检测温度下IL 2活性高于 37℃

Molecular characterization and expression profiles in response to bacterial infection of Chinese soft-shelled turtle interleukin-8 (IL-8), the first reptilian chemokine gene

In this study, an IL-8 homologue has been cloned and identified from a reptile, Chinese soft-shelled turtle for the first time. The full-length cDNA of turtle IL-8 was 1188 bp and contained a 312 bp open reading frame (ORF) coding for a protein of 104 amino acids. The chemokine CXC domain, which contained Glu-Leu-Arg (ELR) motif and four cysteine residues, was well conserved in turtle IL-8. The 4924 bp genomic DNA of turtle IL-8 contained four exons and three introns. Phylogenetic analysis showed that the amino acid sequence of turtle IL-8 clustered together with birds. RT-PCR analysis showed that turtle IL-8 mRNA was constitutively expressed liver, spleen, kidney, heart, blood and intestine tissues of control turtles. Real-time quantitative PCR analysis further indicated that the turtle IL-8 mRNA expression was apparent in various tissues at 8 h and up-regulated significantly during 8 h-7 d after Aeromonas hydrophila infection. The present studies will help us to understand the evolution of IL-8 molecule and the inflammatory response mechanism in reptiles. (C) 2009 Elsevier Ltd. All rights reserved.

Molecular cloning, promoter analysis and induced expression of the complement component C9 gene in the grass carp Ctenopharyngodon idella

Complement-mediated killing of pathogens through lytic pathway is an important effector mechanism of innate immune response. C9 is the ninth member of complement components, creating the membrane attack complex (MAC). In the present study, a putative cDNA sequence encoding the 650 amino acids of C9 and its genomic organization were identified in grass carp Ctenopharyngodon idella. The deduced amino acid sequence of grass carp C9 (gcC9) showed 48% and 38.5% identity to Japanese flounder and human C9, respectively. Domain search revealed that gcC9 contains a LDL receptor domain, an EGF precursor domain, a MACPF domain and two TSP domain located in the N-terminal and C-terminal, respectively. Phylogenetic analysis demonstrated that gcC9 is clustered in a same clade with Japanese flounder, pufferfish and rainbow trout C9. The gcC9 gene consists of 11 exons with 10 introns, spacing over approximately 7 kb of genomic sequence. Analysis of gcC9 promoter region revealed the presence of a TATA box and some putative transcription factor such as C/EBP, HSF, NF-AT, CHOP-C, HNF-3B, GATA-2, IK-2, EVI- 1, AP-1, CP2 and OCT-1 binding sites. The first intron region contains C/EBPb, HFH-1 and Oct-1 binding sites. RT-PCR and Western blotting analysis demonstrated that the mRNA and protein of gcC9 gene have similar expression patterns, being constitutively expressed in all organs examined of healthy fish, with the highest level in hepatopancreas. By real-time quantitative RT-PCR analysis, gcC9 transcripts were significantly up-regulated in head kidney, spleen, hepatopancreas and down-regulated in intestine from inactivated fish bacterial pathogen Flavobacterium columnare-stimulated fish, demonstrating the role of C9 in immune response. (c) 2007 Elsevier B.V. All rights reserved.

alr0117, a two-component histidine kinase gene, is involved in heterocyst development in Anabaena sp PCC 7120

Anabaena sp. PCC; 7120 was mutagenized by transposon Tn5-1087b, generating a mutant whose heterocysts lack the envelope polysaccharide layer. The transposon was located between nucleotides 342 and 343 of alr0117, a 918 bp gene encoding a histidine kinase for a two-component regulatory system. Complementation of the mutant with a DNA fragment containing alr0117 and targeted inactivation of the gene confirmed that alr0117 is involved in heterocyst development. RT-PCR showed that alr0117 was constitutively expressed in the presence or absence of a combined-nitrogen source. hepA and patB, the two genes turned on during wild-type heterocyst development, were no longer activated in an alr0117-null mutant. The two-component signal transduction system involving alr0117 may control the formation of the envelope polysaccharide layer and certain late events essential to the function of heterocysts.

Second-harmonic generation of Bogoliubov excitations in a two-component Bose-Einstein condensate

A second-harmonic generation (SHG) is predicted for the Bogoliubov excitations in a two-component Bose-Einstein condensate. It is shown that, because the linear dispersion curve of the excitations displays two branches, the phase-matching condition for the SHG can be fulfilled if the wave vectors and frequencies of fundamental and second-harmonic waves are selected suitably from different branches. The nonlinearly coupled envelope equations for the SHG are derived by using a method of multiple scales. The explicit solutions of these envelope equations are provided and the conversion efficiency of the SHG is also discussed.

The quantum tunnelling between two-component Bose-Einstein condensates in a double-well configuration

We examine in terms of exact solutions of the time-dependent Schrodinger equation, the quantum tunnelling process in Bose-Einstein condensates of two interacting species trapped in a double well configuration. Based on the two series of time-dependent SU(2) gauge transformations, we diagonalize the Hamilton operator and obtain analytic time-evolution formulas of the population imbalance and the berry phase. the particle population imbalance (a(L)(+)aL - a(R)(+)a(R)) of species A between the two wells is studied analytically.

The quantum tunneling between two-component Bose-Einstein condensates in a double-well configuration

In terms of exact solution of the time-dependent Schrodinger equation. we examine the quantum tunneling process in Bose condensates of two interacting species trapped in a double well configuration. We use the two series of time-dependent SU(2) gauge transformation to diagonalize the Hamilton operator obtain analytic time-evolution formulas of the population imbalance and the berry phase. The particle population imbalance (a(L)(+)a(L) - a(R)(+)a(R)) of species A between the two wells is studied analytically.