7号淀粉酶链霉菌M1033木糖异构酶基因的启动子活性检测,转录起始点…

分别从重组质粒ＰＵＢ１及其Ｍ１３亚克隆Ｓ１中将７号淀粉酶链霉菌（Ｓｔｒｃｐｔｏｍｙｃｅｓ ｄｉａｓｔａｔｉｃｕｓ Ｎｏ．７）Ｍ１０３３（以下简称Ｓ。ｄｉ．Ｍ１０３３）木糖异构酶基因的－１９２－＋５８１ｂｐ片段克隆入链霉菌启动子探测质粒ＰＩＪ４０８３中,转化变铅青链霉菌（Ｓ。ｌｉｖｉｄａｎｓ）ＴＫ２４。通过对其邻苯二酚加双氧酶活性的检测表明,该片段具有启动子活性。应用Ｍ１３亚克隆Ｓ１和合成引物Ｐ６延     

麦迪霉素产生菌中具强启动活性DNA片段的结构与功能分析

用启动子探针质粒ｐＩＪ４８６从麦迪霉素产生菌（Ｓ．ｍｙｃａｒｏｆａｃｉｅｎｓ）中克隆到１个具启动功能的ＨｉｎｄＩＩＩ－ＨｉｎｄＩＩＩ２．０ｋｂ片段,含该片段的重组质粒ｐ４Ｈ２转化子在ＭＭ基本培养基上对卡那霉素（Ｋｍ）抗性可达５００μｇ／ｍｌ以上。亚克隆缺失分析结果表明,该片段不同部分的缺失对启动活性有不同程度的影响,说明它具有较复杂的转录调控机制。ＤＮＡ序列分析结果显示,该片段含有１９８４个核苷酸,其Ｇ＋Ｃ％为４７．７％,不存在典型的链霉菌的可读框;进一步的分析发现,其６５０ｂｐ、１１５０ｂｐ和１５００ｂｐ区域分别与麦迪霉素酮基还原酶基因等链霉菌基因的启动子序列具有同源性;在５２０－５７０ｂｐ区域与大肠杆菌ｔＲＮＡ基因上游激活序列（ＵＡＳ）有较好的同源性,提示链霉菌中可能存在可增强基因转录的ＤＮＡ元件。     

链霉菌质粒pSGL1最小复制子序列测定及分析

质粒ｐＳＧＬ１（７．４ｋｂ）是从球孢链霉菌（Ｓｔｒｅｐｔｏｍｙｃｅｓｇｌｏｂｉｓｐｏｒｕｓ）中分离得到的一个高拷贝质粒,已证明其最小复制子位于Ｓａｕ３ＡＩ酶切的２０ｋｂ片段上。对该片段进行亚克隆,测序后数据表明该片段是一个新序列。仅有一个开放阅读框架（ＯＲＦＲ）位于最小复制子中,推测其编码的蛋白质含有滚环复制质粒复制酶的特定序列。     

链霉菌—大肠菌穿梭质袜载体pSGLgpp的构建及应用

质粒ｐＳＧＬ１（７．４ｋｂ）是从球孢链霉菌（Ｓｔｒｅｐｔｏｍｙｃｅｓ ｇｅｏｂｉｓｐｏｒｕｓ）中分离得到的一个高拷贝质粒,已测定其最小复制子序列。从球孢链霉菌总ＤＮＡ中采用ＰＣＲ方法扩增获得编码Ｃ１０２７前蛋白信号肽的ＤＮＡ片断ｇｐｐ。将ｇｐｐ克隆至ｐＳＧＬ１的衍生质粒ｐＳＧＬＮ中,获得新的链霉菌表达型质粒载体ｐＳＧＬｇｐｐ。应用该质粒进行了人的河溶性白细胞介素１受体Ｉ型的表达。     

Mg~(2+)、Co~(2+)、Mn~(2+)和Ca~(2+)对葡萄糖异构酶活性的影响

Ｄ－木糖异构酶（ＥＣ５．３．１．５）能催化醛糖Ｄ－木糖至酮糖Ｄ－木酮糖的转化,又能催化Ｄ－葡萄糖至Ｄ－果糖的异构化反应．故被称为葡萄糖异构酶（ＧＩ）．一般认为,葡萄糖异构酶存在二种作用机制：烯醇化和氢转移机制．对我国自行筛选的链霉菌Ｎｏ．７Ｍ１０３３菌株葡萄糖异构酶的晶体结构研究表明［１］,异构化可能是采用氢转移机制．这一机制认为催化需要半径≤０．０８ｎｍ的二价金属离子如Ｍｇ２＋、Ｃｏ２＋和Ｍｎ２＋等激活,而离子半径＞０．０８ｎｍ的二价金属离子则对酶的活性有抑制作用．Ｍｇ２＋、Ｃｏ２＋和Ｍｎ２＋…     

日本血吸虫26kD抗原基因在BCG中的表达

研究了外源基因日本血吸虫２６ｋＤ抗原（Ｓｊ２６ＧＳＴ）在卡介苗（ｂａｃｉｌｕｓＣａｌｍｅｔｅ－Ｇｕｅｒｉｎ,ＢＣＧ）、耻垢分枝杆菌（Ｍ．ｓｍｅｇｍａｔｉｓ）和大肠杆菌（Ｅ．ｃｏｌｉ）中的表达．运用重组ＤＮＡ和聚合酶链反应（ＰＣＲ）等分子生物学技术,以表达Ｓｊ２６ＧＳＴ的Ｅ．ｃｏｌｉｐＧＥＸ衍生质粒为模板,经ＰＣＲ得到编码Ｓｊ２６ＧＳＴ的全长ｃＤＮＡ片段．将其按正确的阅读框顺序,克隆到人结核杆菌热休克蛋白（ｈｅａｔｓｈｏｃｋｐｒｏｔｅｉｎ,ＨＳＰ）７０的启动子下游,再将ＨＳＰ７０启动子和Ｓｊ２６ＧＳＴ基因一起亚克隆到Ｅ．ｃｏｌｉ－分枝杆菌穿梭质粒ｐＢＣＧ－２０００中,得到Ｅ．ｃｏｌｉ－分枝杆菌穿梭表达质粒ｐＢＣＧ－Ｓｊ２６．ｐＢＣＧ－Ｓｊ２６电转化入ＢＣＧ和Ｍ．ｓｍｅｇｍａｔｉｓｍｃ２１５５中表达Ｓｊ２６ＧＳＴ抗原,所表达的天然重组Ｓｊ２６ＧＳＴ（ｒＳｊ２６ＧＳＴ）为可溶性蛋白,在ＳＤＳ－ＰＡＧＥ上分子量为２６ｋＤ处可见明显的表达蛋白带．其表达量分别占ＢＣＧ和Ｍ．ｓｍｅｇｍａｔｉｓ菌体总蛋白的１５％和１０％．可见,Ｓｊ２６ＧＳＴ基因能在ＢＣＧ中高效表达．     

变性双链法实现葡萄糖异构酶基因敲除的研究

对７号淀粉酶菌Ｍ１０３３的遗传背景进行分析,建立了其原生质体制备、转化的优化条件。构建了葡萄糖异构酶（ＧＩ）结构基因内插千方百计以链丝菌肽基因（ｔｓｒ）的置换型同源重组质粒,利用其变性双链片段实现与Ｍ１０３３菌株染色体上基因的同源重组,获得置换型葡萄异构酶缺陷型蓖Ｍ１０３３ＬＪ。为在染色体上引入突变位点实现染色体上分子定点改造造尊定了基础。     

人肿瘤坏死因子基因在变铅青链霉菌中的克隆与表达

以质粒ｐＩＪ４８６为载体,将来源于质粒ｐＨＴ１的肿瘤坏死坏因子（ＴＮＦ－ａ）ｃＤＮＡ克隆至变铅青链霉菌,以新霉素（３０μｇ／ｍｌ）为选择标记,获得了数百转化子,实验表明Ｎｏ．７转化子ｓ．ｌｉｖｉｄａｎｓ ＴＫ５４－ＨＴ所含重组质粒ｐＩＪＴ７已克隆有ＴＮＦｃＤＮＡ。Ｌ９２９细胞毒实验结果表明该转化子ＴＮＦ表达量可达１０＾８活性单位／升以上,中和实验确证其表达产物为人ＴＮＦ－ａ,ＳＤＳ－ＰＡＧＥ表明克     

牛疱疹病毒I型前早期基因BICPO的表达对牛免疫缺陷病毒LTR的反式激活…

由含有ＢＨＶ－１（Ｂｏｖｉｎｅ Ｈｅｒｐｅｓ Ｖｉｒｕｓ－１）前早期基因的基因组片段亚克隆ＩＣＰＯ（ＢＨＶ－１Ｉｎｆｅｃｔｅｄ Ｃｅｌｌ Ｐｒｏｔｅｉｎ Ｏ）的ＤＮＡ序列至表达载体ｐＳＶＤ３,构建质粒ｐＳＶ２．９。将该质粒与ＰＢＬＴＲ－Ｌｕｃ共转染小牛肺细胞,检测转染细胞裂解物的荧光素酶活性,ＢＩＣＰＯ的表达产物可以显著地激活ＢＩＶ ＬＴＲ启动子控制下的荧光素酶基因的表达。根据ＰＳＶ２．９与含有Ｂ     

长果升麻的化学成分研究

从长果升麻（Ｓｏｕｌｉｅａ ｖａｇｉｎａｔａ）根茎中分离出６种皂甙,经光谱（ＦＡＢ－ＭＳ、１Ｈ－ＮＭＲ、１Ｈ－１Ｈ－ＣＯＳＹ、１３Ｃ－ＮＭＲ、１Ｈ－１３Ｃ－ＣＯＳＹ）分析,分别鉴定为２７－ｄｅｏｘｙａｃｔｅｉｎ（１）,ａｃｔｅｉｎ（２）,２５－０－乙酰升麻醇木糖甙（３）,２５－甲基升麻醇木糖甙（４）,升麻醇木糖甙（５）,２４－ａｃｅｔｙｌｈｙｄｒｏｓｈｅｎｇｍａｎｏｌ ｘｙｌｏｓｉｄｅ（６）,其中     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.