草鱼(Ctenopharyngodon idellus)线粒体DNA控制区及其两侧tRNA基因的克隆与结构分析

动物线粒体ＤＮＡ控制区是线粒体基因组复制与基因表达的最主要的调控区．采用杂交和测序的方法对草鱼线粒体ＤＮＡ控制区进行定位、克隆并测定了控制区及其旁侧的ｔＲＮＡＰｈｅ、ｒＲＮＡＰｒｏ和ｒＲＮＡＴｈｒ三个基因的序列,与多种脊椎动物的相应序列进行了比较,并进行了结构分析．草鱼线粒体控制区全长９２７ｂｐ,含有与酵母和爪蟾线粒体启动子相似的序列,其ＣＳＢⅠ、ＣＳＢⅡ和ＣＳＢⅢ序列与其他几种动物的ＣＳＢ比较相当保守,ＴＡＳ与其回文基序可形成稳定的茎环结构,成为Ｈ－链复制的终止信号．草鱼线粒体ｔＲＮＡＰｈｅ、ｔＲＮＡＰｒｏ和ｔＲＮＡＴｈｒ可折叠成三叶草形二级结构,其基因具有许多不同于细胞质ｔＲＮＡ基因的结构特点,可能反映了线粒体ｔＲＮＡ与线粒体核糖体具有不寻常的作用方式     

Pollen transfer dynamics and the evolution of gametophytic self-incompatibility

Recent studies of mating system evolution have attempted to include aspects of pollination biology in analysis of both theoretical models and experimental systems. In light of this growing trend, we propose a simple population genetic model for the evolution of gametophytic self-incompatibility, incorporating parameters for pollen discounting and pollen export/capture. In this model, we consider several cases that span the spectrum for dominance of the mutant self-incompatibility allele and for the degree of incompatibility conferred by the allele. We confirm earlier results that inbreeding depression is required for successful invasion of the self-incompatibility allele and we demonstrate that, unless pollen discounting is very low, the level of inbreeding depression must be very high for an allele conferring self-incompatibility to become established. Finally, we show that the dominance of the mutant allele has a greater impact on the fate of a newly arisen self-incompatibility allele than the strength of the incompatibility conferred by the allele. In particular, the more recessive the self-incompatibility expression in heterozygote stigmas and the weaker the response induced, the easier it is for a self-incompatibility allele to invade.     

开封沙地试验区不同作物种植方式的综合效益评价

开封沙区地处河南省沙区腹地,风沙灾害严重,旱涝灾害频繁,地力瘠薄,严重障碍着沙区农牧业生产的发展。因此,自１９８３年在国家科技攻关的基础上建立了开封沙地试验区,探索平原沙区农业生产的发展模式。尤其是“八五”攻关期间,深化农林牧复合生态系统的优化研究,...     

Rapid bidirectional modulation of mRNA expression and export accompany long‐term facilitation and depression of Aplysia synapses

Serotonin (5‐HT) and the neuropeptide Phe‐Met‐Arg‐Phe‐amide (FMRFa) modulate synaptic efficacy of sensory neurons (SNs) of Aplysia in opposite directions and for long duration. Both long‐term responses require changes in mRNA and protein synthesis. The SN‐specific neuropeptide, sensorin A, is a gene product that appears to be increased by 5‐HT and decreased by FMRFa. We examined whether changes in sensorin A mRNA levels in the cell body and neurites of SNs accompany long‐term facilitation and depression. Both 5‐HT and FMRFa evoked rapid changes in sensorin A mRNA levels in the SN cell bodies: an increase with 5‐HT and a decrease with FMRFa. Parallel changes in sensorin A mRNA levels in SN neurites were detected 2 h and 4 h later. These rapid changes in mRNA expression and net export required the presence of the appropriate target motor cell L7. The neuromodulators failed to produce changes in mRNA expression or export when SNs were cultured alone or with the inappropriate target cell L11. The changes in mRNA expression were transient because mRNA levels returned to control values 24 h after treatment, while synaptic efficacy remained altered by the respective treatments. These results indicate that two neuromodulators produce distinct, but transient, target‐dependent effects on expression and export of a cell‐specific mRNA that correlate with changes in synaptic plasticity. © 2000 John Wiley & Sons, Inc. J Neurobiol 46: 41–47, 2001     

The protease-sensitive N-terminal polybasic region of prion protein modulates its conversion to the pathogenic prion conformer

Conversion of normal prion protein (PrP^C) to the pathogenic PrP^Sc conformer is central to prion diseases such as Creutzfeldt–Jakob disease and scrapie; however, the detailed mechanism of this conversion remains obscure. To investigate how the N-terminal polybasic region of PrP (NPR) influences the PrP^C-to-PrP^Sc conversion, we analyzed two PrP mutants: ΔN6 (deletion of all six amino acids in NPR) and Met4-1 (replacement of four positively charged amino acids in NPR with methionine). We found that ΔN6 and Met4-1 differentially impacted the binding of recombinant PrP (recPrP) to the negatively charged phospholipid 1-palmitoyl-2-oleoylphosphatidylglycerol, a nonprotein cofactor that facilitates PrP conversion. Both mutant recPrPs were able to form recombinant prion (recPrP^Sc) in vitro, but the convertibility was greatly reduced, with ΔN6 displaying the lowest convertibility. Prion infection assays in mammalian RK13 cells expressing WT or NPR-mutant PrPs confirmed these differences in convertibility, indicating that the NPR affects the conversion of both bacterially expressed recPrP and post-translationally modified PrP in eukaryotic cells. We also found that both WT and mutant recPrP^Sc conformers caused prion disease in WT mice with a 100% attack rate, but the incubation times and neuropathological changes caused by two recPrP^Sc mutants were significantly different from each other and from that of WT recPrP^Sc. Together, our results support that the NPR greatly influences PrP^C-to-PrP^Sc conversion, but it is not essential for the generation of PrP^Sc. Moreover, the significant differences between ΔN6 and Met4-1 suggest that not only charge but also the identity of amino acids in NPR is important to PrP conversion.     

Chemical synthesis of human protein S thrombin-sensitive module and first epidermal growth factor module

Human plasma protein S is a nonenzymatic cofactor for activated protein C (APC) in the inactivation of coagulation factors Va and VIIIa, and helps to provide an essential negative feedback on blood coagulation. Previous indirect evidence suggested that the thrombin-sensitive region (TSR:residues 47–75, 1 disulfide) and the first epidermal growth factorlike region (EGF1: residues 76–116, 3 disulfides) of protein S may be functionally important for expression of its APC cofactor activity. To study the functional importance of these modules directly, access to the isolated TSR and EGF1 modules would be preferred. Recombinant expression of protein S intact TSR and correctly folded EGF1 has not been possible. Here we describe the synthesis of both TSR and EGF1 modules by stepwise solid phase peptide synthesis using the in situ neutralization/2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate activation procedure for tert-butoxycarbonyl chemistry. For the TSR, correct intramodular disulfide bonding was confirmed. To overcome folding difficulties with the EGF1, a two-step oxidation procedure was used in which the cysteines involved in the middle, crossing, disulfide bond (Cys⁸⁵-Cys¹⁰²) remained protected with acetamidomethyl (Acm) groups after hydrogen fluoride treatment of the peptide resin. Selective formation of the first two disulfide bonds (Cys⁸⁰-Cys⁹³ and Cys¹⁰⁴-Cys¹¹³) was followed by release of the Acm groups and subsequent formation of the third disulfide bond (Cys⁸⁵-Cys¹⁰²). CD studies revealed 54% of β-sheet/turn in the EGF1 that is characteristic for EGF modules. Deuterium exchange studies suggested a very tightly packed core in EGF1 that is not accessible to the bulk solvent, likely a result from the compact structure caused by its three disulfide bonds. The 30% β-sheet structure observed in the TSR involved amide protons that could be readily exchanged by deuterons, likely reflecting a more flexible structure of the TSR loop in contrast to the rigid structure of EGF1. The establishment of synthetic access to the TSR and EGF1 of protein S provides a versatile tool to study interactions of these modules with the blood coagulation components of the anticoagulant plasma protein C pathway. © 1998 John Wiley & Sons, Inc. Biopoly 46: 53–63, 1998     

mtDNA and language support a common origin of Micronesians and Polynesians in Island Southeast Asia

The origins and relationships among Micronesians, Polynesians, and Melanesians were investigated. Five different mtDNA region V length polymorphisms from 873 individuals representing 24 Oceanic and Asian populations were analyzed. The frequency cline of a common deletion and the distributions of a rare expanded length polymorphism support the origin of both Micronesians and Polynesians in Island Southeast Asia. Genetic, linguistic, and geographic distances were compared to assess the relative importance of isolation and gene flow during the prehistory of 19 Austronesian-speaking populations subdivided into five potential spheres of interaction. We observed significant correlations (P < 0.05) between genetic and linguistic distances in four of five comparisons. These data indicate extensive gene flow throughout much of Micronesia, but substantial isolation in other Pacific regions. Although recent advancements in our understanding of intentional voyaging within Remote Oceania have challenged the existence of the “myth of the primitive isolate,” we caution against the adoption of panmictic alternatives. Am J Phys Anthropol 105:109–119, 1998. © 1998 Wiley-Liss, Inc.     

Identification of cyanobacteria by polymorphisms of PCR-amplified ribosomal DNA spacer region

The 16S-23S ribosomal DNA spacer region of selected cyanobacterial strains was amplified by the polymerase chain reaction using primers to conserved flanking sequences. Single or multiple rDNA amplification products were generated depending on the strain and primer pair. Species could generally be distinguished on the basis of size heterogeneity of the products. Analysis of restriction digests of the amplified rDNAs indicated polymorphisms useful in identification. Four enzymes (HinfI, DdeI, AluI, TaqI) generated restriction fragment length patterns that could discriminate between the cyanobacteria to the taxonomic levels of genus and species. This approach should prove useful in the rapid identification of cyanobacteria.     

Mitochondrial DNA variation in Koryaks and Itel'men: Population replacement in the Okhotsk Sea–Bering Sea region during the neolithic

In this study, we analyzed the mitochondrial DNA (mtDNA) variation in 202 individuals representing one Itel'men and three Koryak populations from different parts of the Kamchatka peninsula. All mtDNAs were subjected to high resolution restriction (RFLP) analysis and control region (CR) sequencing, and the resulting data were combined with those available for other Siberian and east Asian populations and subjected to statistical and phylogenetic analysis. Together, the Koryaks and Itel'men were found to have mtDNAs belonging to three (A, C, and D) of the four major haplotype groups (haplogroups) observed in Siberian and Native American populations (A–D). In addition, they exhibited mtDNAs belonging to haplogroups G, Y, and Z, which were formerly called “Other” mtDNAs. While Kamchatka harbored the highest frequencies of haplogroup G mtDNAs, which were widely distributed in eastern Siberian and adjacent east Asian populations, the distribution of haplogroup Y was restricted within a relatively small area and pointed to the lower Amur River–Sakhalin Island region as its place of origin. In contrast, the pattern of distribution and the origin of haplogroup Z mtDNAs remained unclear. Furthermore, phylogenetic and statistical analyses showed that Koryaks and Itel'men had stronger genetic affinities with eastern Siberian/east Asian populations than to those of the north Pacific Rim. These results were consistent with colonization events associated with the relatively recent immigration to Kamchatka of new tribes from the Siberian mainland region, although remnants of ancient Beringian populations were still evident in the Koryak and Itel'men gene pools. Am J Phys Anthropol 108:1–39, 1999. © 1999 Wiley‐Liss, Inc.