Differential effects of the serotonin receptors on cultured rat cerebral cortical neurons

Effects of serotonin (5-HT) on cerebral cortical neurons were examined by patch clamp techniques. 5-HT produced a variety of responses such as outward (19/73 patches/neurons), slow inward (15/73 patches/neurons), fast inward (8/73 patches/neurons), and mixed currents (initially fast inward deflection followed by an outward response: 2/73 patches/neurons), with a latency of 12 sec, 15 sec, 0 sec, and 0 sec respectively, at a holding potential of −60 mV in whole-cell patches. The fast inward currents were again evoked by a selective 5-HT3 receptor agonist, 1-(m-chlorophenyl)-biguanide hydrochloride (CPBG). In the cell-attached patch clamp configuration, 5-HT inside the patch pipette elicited single channel currents with slope conductances of 42 pS and 132 pS (4/42 patches/neurons). CPBG inside the patch pipette evoked inward single channel currents with a lower slope conductance of 41 pS (3/23 patches/neurons). In contrast, application of 5-HT or a 5-HT₂ receptor agonist, α-methyl-5-hydroxytryptamine-maleate, outside the patch pipette induced outward single channel currents with a major slope conductance of 140 pS (8/30 patches/neurons) or 135 pS (6/20 patches/neurons), respectively. These results indicate that the outward and fast inward currents may be mediated respectively by the 5-HT₂ receptor, which is coupled to a G-protein, and by the 5-HT₃ receptor, which contains the non-selective cation channel, and that the mixed type may be caused by both the 5-HT₂ and 5-HT₃ receptors. Received 27 September 1996; received after revision 4 November 1996; accepted 7 November 1996     

把基因分析引进人类学——人类学学者访谈录之三十九

结合自身的研究,探讨了遗传学在人类学的应用及前景问题,如体质人类学、医学人类学、分子人类学、考古人类学等分支领域。     

Strategy for the mapping of interactive genes using bulked segregant analysis method and Mapmaker/Exp software

A qualitative trait is usually controlled by a single gene, but it may be sometimes controlled by two or even more genes. This phenomenon is called gene interaction. Rapidly searching for linked mo- lecular markers via bulked segregant analysis (BSA) and then constructing regional linkage map with Mapmaker/Exp has become a common approach to mapping single major genes. However, methods and computer programs developed for mapping single major genes cannot be simply applied to interactive genes because the genetic patterns of gene interac- tions are quite different from that of single-gene in- heritance. Up to now, experimental methods for quickly screening molecular markers linked to inter- active genes and statistical methods and corre- sponding computer softwares for simultaneously analyzing the linkage relationships of multiple mo- lecular markers to an interactive gene have not been available. To solve this problem, in this paper, we propose a strategy for mapping interactive genes using BSA and Mapmaker/Exp. We demonstrate that all interactive genes can be mapped by the 'BSA Mapmaker/Exp' strategy using F2 generation (in a few cases, F3 generation is also needed). As BSA and Mapmaker/Exp have been broadly used in gene mapping studies and are well known by many re- searchers, the strategies proposed in this paper will be useful for practical researches.     

以mRNA差异显示法分离牙鲆白化相关基因的初步研究

以正常和白化牙鲆表皮组织总RNA为模板,用Oligo d(T)13M(M分别为A、C、G)3种锚定引物和26种差异显示随机引物组合进行差异显示,PAGE凝胶电泳后,用新型高灵敏度核酸荧光染料SYBR GREEN I显示差异DNA条带,得到区分度较好的差异表达图谱,共分离49条牙鲆白化相关DNA片段,片段长度260～800bp左右,经克隆测序后将可作为新的ESTs进行同源序列比较.     

基因组研究离诠释生命奥妙还有多远

基因组研究表明，人类与黑猩猩和老鼠的基因序列98％以上是相同的，是否是这1％～2％的基因的差异，就决定了人与黑猩猩和老鼠在表现型方面很大的差异?本文通过大量分析表明，由碱基(5个)→核酸[DNA→RNA(mRNA、rRNA、tRNA)]→氨基酸(22种)→蛋白质(数百千种)→染色体→基因组→性状，其遗传变异信息有逐级放大的过程。这是生物间基因序列虽然有98％甚至99％的相同，但表现型差异却较大的原因所在。基因序列完全相同，并不能说明基因功能相同，基因组研究的结果不能完全诠释基因功能表达的复杂过程。今后必将由序列基因组学→结构基因组学→功能基因组学→蛋白质组学深入发展，必须从网络层次研究基因表达，才能揭示生命的奥妙。另外我们使用的各种基因组研究技术和软件还有待改进，才能真正反映生物基因的差异。     

水稻RSSG8基因启动子与GUS融合基因的构建及在烟草中的瞬间表达

使用染色体步移(Genome walking)法，从籼稻(Oryza sativa subsp．indica)桂朝2号基因组中克隆到长度为471bp的水稻精细胞优势表达基因RSSG8的启动子片段R8PN，并进行了全序列测定和分析．该段序列中含有3个CAAT-box，6个Gobox和多种植物顺式作用元件，但没有发现典型的TATA-box，推测为一种特殊的启动子结构，为了鉴定RSSG8基因的基本启动子元件，将二条长度不同的5’端侧翼区缺失体(分别长471bp，260bp)定向插入载体pBI121中，取代原有的CaMV 35S启动子，构了驱动报告基因GUS的植物表达载体pRGUS1，pRGUS2，通过农杆菌介导的瞬时表达法转化烟草叶片和花粉，快速鉴定启动子片段中起关键作用的区域，结果显示两个缺失片段都能启动GUS的表达，可以初步判定这两个片段具有启动子功能。     

全长基因的克隆

新基因全长cDNA序列、全长DNA序列的获得常常是分子生物学工作者面临的难题。本文就克隆全长基因的各种技术如文库筛选法、各种PCR技术及新发展起来的电子克隆法等方法作一简单综述。     

地衣芽孢杆菌A13重组表达黑曲霉植酸酶phyA基因的研究

将黑曲霉Z6植酸酶phyA基因置于芽孢杆菌强启动子F1下,与大肠-芽孢杆菌穿梭表达载体pHY300PLK连接,重组质粒pHY300-F1-phyA电击转化地衣芽孢杆菌A13。筛选阳性克隆,获得重组菌株A13-F1-phyA,PCR扩增及酶切验证表明植酸酶基因已转入地衣芽孢杆菌。SDS-PAGE分析和表达产物的酶活性研究证明,重组菌株中植酸酶得到了有效分泌和表达。     

生态风险评价

本文阐述了生态风险评价的基本概念和定量研究方法．通过研究方法与技术路线,确立评价要素和受体—端点,建立一套风险评价的生态系统指标体系