Site-specific cleavage of chromosomes in vitro through Cre-lox recombination.

Site-specific recombination systems are useful tools for chromosome engineering in vivo and site-specific DNA cleavage methods have applications in genome analysis and gene isolation. Here, we report a new method to fragment chromosomes in vitro using the Cre-lox site-specific recombination system. Two lox sites were targeted into the 5.7 Mb chromosomes I of Schizosaccharomyces pombe. In vitro recombination between chromosomal lox sites and exogenously provided lox oligonucleotides 'cleaved' the chromosome at the defined lox sequences. Site-specific cleavage of lox sites in the tobacco genome was also demonstrated. This recombination-based cleavage method provides a novel approach for structural and functional analyses of eukaryotic chromosomes as it allows direct isolation of chromosome regions that correspond to phenotypes revealed through Cre-lox mediated chromosome rearrangements in vivo. Moreover, recombination with end-labeled lox oligonucleotides would permit the specific end-labeling of chromosome segments to facilitate the long range mapping of chromosomes.     

Distribution of 3α-hydroxysteroid dehydrogenase in rat brain and molecular cloning of multiple cDNAs encoding structurally related proteins in humans

3α-Hydroxysteroid dehydrogenase in the brain is responsible for production of neuroactive tetrahydrosteroids that interact with the major inhibitory gamma-aminobutyric acid receptor complexes. Distribution of 3α-hydroxysteroid dehydrogenase in different regions of the brain in rats was evaluated by activity assay and by Western immunoblotting using a monoclonal antibody against liver 3α-hydroxysteroid dehydrogenase as the probe. The olfactory bulb was found to contain the highest level of 3α-hydroxysteroid dehydrogenase activity, while moderate levels of the enzyme activity were found in other regions such as cerebellum, cerebral cortex, hypothalamus and pituitary. Some activity was found in the rest of the brain such as amygdala, brain stem, caudate putamen, cingulate cortex, hippocampus, midbrain, and thalamus. The protein levels of 3α-hydroxysteroid dehydrogenase in different regions of the brain as detected by Western immunoblotting are comparable to those of the enzyme activity. We used the rat cDNA as the probe to screen a human liver λ gt11 cDNA library. A total of four different cDNAs were identified and sequenced. One of the cDNAs is identical to that of the human chlordecone reductase cDNA except that our clone contains a much longer 5′-coding sequence than previously reported. The other three cDNAs display high degrees of sequence homology to those of both rat 3α-hydroxysteroid dehydrogenase and human chlordecone reductase. We are currently investigating the functional relationship between the enzymes encoded by these human cDNAs and 3α-hydroxysteroid dehydrogenase.     

细胞周期和调控因子

近期对细胞周期调控的研究获得了突破性进展.人们深入地研究了周期蛋白家族和pp³⁴蛋白家族在周期调控中的作用及二者之间的相互关系,同时还发现了很多与之相关的调控因子,它们彼此相互作用,形成了极为复杂的级联调控网络.     

昆虫神经毒素的研究:各种神经毒剂引起毒素的产生

用五类(七种)神经毒性的杀虫药剂,对美洲(虫非)蠊测定了它们能否引起血淋巴中毒素的产生。DDT、溴氰菊酯及六六六均能引起血淋巴中毒素的产生,而E605、西维因、巴丹及杀虫脒均无此效果。纸层析及薄层层析,用标准样品(酪氨酸、酪胺、苯乙胺、L-亮氨酸及异戊胺)作比较,测定了这一产生的血淋巴中的毒素乃是酪胺,或主要是酪胺。增效试验,证明了杀虫脒与DDT或溴氰菊酯合用时,能增加酪胺的产量。讨论了这一毒素产生的条件,以及这一毒素有可能不是单一成分,而是几种单胺及氨基酸的复合物,但酪胺为其主要成分。     

我国西部地区蚤目四种未知雌性的记述

本文系青海省蚤目研究的第四报。首次记述蚤目的四种未知雌性。雄性根据我国西藏采获的标本已先后有人记述:近缘纤蚤Rhadinopsylla accola Wagner,1930;端圆盖蚤Callopsylla kozlovi(Wagner.1929);卷带倍蚤Amphalius sptrataenius Liu,Wu ＆ Wu,1966;短突角叶蚤Ceratophyllus breviprolectus Liu,Wu ＆ Wu,1966。     

Mycotoxin Producing Potential of Fungi Associated with Dry Shrimps

Among 114 fungi isolated from 20 dry shrimp samples, 27 isolates were capable of producing mycotoxins. Aspergillus and Penicillium were-the-predominant fungi. Two of 3 Aspergillus flavus could produce aflatoxin B₁ and G₁ on yeast extract sucrose medium and on surface sterilized dry shrimps. For other toxigenic species, toxicity was found mostly in the chloroform extracts of cultures. Visibly mouldy dry shrimps had significantly higher moisture content than the non-mouldy ones. Most of the fungi were isolated from mouldy samples.     

性信息素对红铃虫雄蛾近距离性行为的抑制效应

鳞翅目昆虫雄蛾在交配之前的性行为可以分为远距离对雌蛾的定向和包括着陆、搜索、交配等直接导致雌、雄蛾交配成功的近距离行为。这两类不同的行为可由复合性信息素中的不同化学成分引起(Roelofs et al.,1977)或者由同种物质不同浓度所诱发,即雄蛾接受较低浓度刺激时出现远距离定向飞行;而高浓度诱发雄蛾近距离的交配性行为反应(Bartell et al.,1969)。从棉花红铃虫[Pectinophora gossypiella(Saund.)]雌蛾体内已经鉴定得到两种化学成份,顺-7,顺-11-十六碳二烯醇醋酸酯和顺-7,反-11-十六碳二烯醇醋酸酯。这两种成份的1:1混合物称为红铃虫性诱素(Gossyplure)(Hummel et al.,1973),它具有在田间引诱红铃虫雄蛾和诱发雄蛾近距离行为反应的作用。 在田间释放性信息化合物来扰乱雌、雄蛾之间的性信息联系,从而抑制其交配和繁殖是一种有希望的害虫防治方法。用红铃虫性诱素扰乱红铃虫交配的田间试验已取得一定成效(Gaston et al.,1977)。但是我们在田间试验中发现,性诱素抑制红铃虫雄蛾远距离定向后,田间还有相当比例的雌蛾发生了交配,这个现象表明,性诱素还未能完全抑制红铃虫雄蛾的近距离行为。为此我们进行了本项试验,用以阐明环境中红铃虫性诱素的存在对雄蛾近距离性行为的影响。     

乙型脑炎病毒NS2A-C60A位点突变对其生物学特性影响研究*

目的:旨在探索Ⅰ型日本乙型脑炎病毒传代致弱后基因组突变NS2A-C60A对乙脑病毒生物学特性的影响。方法:首先通过对传代致弱及原始乙脑毒株基因组序列进行测序比对、结构预测分析并利用Western blotting(WB)确定了目标研究位点NS2A-C60A;然后使用反向遗传定点突变技术构建拯救了包含NS2A-C60A单点突变的病毒株;最后利用噬斑形态观察、生长曲线、双萤光素酶分析,WB以及炎性因子检测和动物实验研究了该单点突变对于乙脑病毒生物学特性的影响。结果:首次研究发现Ⅰ型乙脑病毒传代致弱会导致NS1'蛋白表达的显著下降以及可能的相关位点NS2A-C60A,并成功拯救获得了NS2A-C60A单点突变毒株rJEV-C60A,研究发现NS2A-C60A突变对乙脑病毒的生长特性及噬斑形成没有显著影响,但是能够显著降低乙脑病毒NS1'蛋白的表达,并且该位点突变能够轻微阻碍乙脑病毒对细胞炎性因子表达的抑制,动物实验结果显示NS2A-C60A点突变病毒与原毒株具有相似的神经毒力,说明该位点突变不是影响乙脑病毒毒力致弱的关键位点。结论:新发现的NS2A-C60A位点突变能够显著减少乙脑病毒NS1'蛋白的表达,但是对其增殖、诱导炎症及神经毒力等生物学特性没有显著影响。     

竹链蚧属一新种记述(同翅目:蚧总科)

在皖南山区调查竹类蚧虫,发现竹链蚧属一新种,现描述如下。模式标本保存于安徽省马鞍山市科委业务科。 竹秆红链蚧Bambusaspia rutilan新种 雌成虫 虫体近卵形,深红或紫红色;长约0.8毫米,宽0.45毫米。缘8字腺一列,后     

人参对大鼠胃G细胞、D细胞影响的免疫组织化学观察

本文应用免疫组织化学PAP方法观察大鼠胃G细胞、D细胞的分布和形态特征,以及人参对胃G细胞、D细胞免疫细胞化学活性的影响。用细胞显微分光光度计检测了服用人参的G细胞、D细胞免疫反应阳性面积及光密度。检测结果表明,人参能使大鼠胃G细胞、D细胞增大,增加G细胞中胃泌素的含量和D细胞中生长抑素的含量。本文的结果提示,人参对大鼠胃G细胞和D细胞的形态及分泌活动有调节性影响。