薄荷伪造桥虫多角体病毒及其核酸的限制性内切酶酶解分析

本文描述了从薄荷伪造桥虫(Argrogramma agnata stgr.)幼虫分离到的一种核型多角体病毒的形态以及采用简便的方法提取的核酸,经限制性内切酶 EcoRI,BamHI,HindⅢ,BglⅠ,BglⅡ和BglⅠ+BglⅡ,BamHl+EcoRl 酶解,获得该病毒核酸的酶解带谱。以入 DNA 的 EcoRl 酶解片段在凝胶中的迁移率与相应 DNA 片段分子量的对数值作标准曲线。从曲线上求得薄荷伪造桥虫多角体病毒核酸酶解各片段的分子量.此病毒核酸的平均分子量为108.51×10~6道尔顿。     

滑鼠蛇肝线粒体DNA的限制酶图谱

用六种限制性内切酶BamHⅠ、EcoRⅠ、PstⅠ、BglⅠ、BglⅡ和SalⅠ对滑鼠蛇肝脏线粒体DNA(mtDNA)进行酶解。发现BglⅡ、PstⅠ、BamHⅠ、BglⅠ和EcoRⅠ在滑鼠蛇肝mtDNA上分别有1、2、3、3和4个切点。SalⅠ不能切割滑鼠蛇肝mtDNA。根据滑鼠蛇肝mtDNA的单酶、双酶完全酶解及部分酶解片段的数目和分子量,建立了滑鼠蛇肝mtDNA的限制酶图谱。     

棉铃虫核多角体病毒DNA的限制性内切酶酶解图谱和电镜观察

棉铃虫核多角体病毒(NPV)加入0.01 Mna₂CO₃—0.05 MNaCl溶解后,用水饱和苯酚-SDS提取NPV DNA。电镜观察证明NPV DNA为非均一性环状结构分子。测量了28个分子的长度,其中15个分子长度为27±5μm,相当分子量为(73±13)×10⁶道尔顿,其余的是较大和较小的分子。限制性内切酶XbaⅠ,XhoⅠ BamHⅠ,EcoRⅠ,BgⅠ Ⅱ分别把NPV DNA酶解为20,9,10,18和11个限制性片段。由片段总和法计算得平均分子量是73.3×10⁶道尔顿。     

三种核型多角体病毒核酸同源性的研究

用分子杂交技术研究了黑点银纹夜蛾(Arqyrogramma agnata Stgr,)单粒包埋型核型多角体病毒(简称A,A,SNPV),斜纹夜蛾(Prodenia litura)多粒包埋型核型多角体病毒(简称PL MNPV)以及用A,A,SNPV感染斜纹夜蛾幼虫所获得的多粒包埋型核型多角体病毒(暂称PoI MNPV)三种病毒核酸的同源性,用SDS-Tris酚法分别提取病毒核酸,用内切酶Eco RⅠ酶解,比较了病毒核酸的酶解图谱及分子量,用〔α-~(32)P〕dATP标记的三种病毒核酸Eco RⅠ酶解片段作探针,分别与各病毒核酸的Eco RI酶解片段杂交,结果表明PoI MNPV与PL MNPV的病毒核酸同源,而A,A,SNPV DNA不与PoI MNPV DNA、PL MNPV DNA杂交,无同源序列。     

转座子Tn233(CH)分子量的测定

转座子Tn233(CH)带有str sul抗性基因,最早是在痢疾杆菌的抗药质粒DR233(Tc~r Cm~r Sm~r Su~r)中发现的。现在通过菌株间的配对,将插入了Tn233(CH)转座子的质粒R144drd3::Tn233(CH)转移到E·coli C600/pBR322(Ap~r、Tc~r)细胞中,组成两种质粒共存的菌株。从此菌株中提取出质粒DNA,用转化方法使它转移到E.coli C600菌株,再从所得到的转化子中用复印方法筛选出Tn233(CH)转座到pBR322质粒的转化子E.coli C600/pBR322::Tn233(CH),然后提出此质粒DNA,经限制性内切酶BamHⅠ、EcoRⅠ、PstⅠ、HindⅢ与PvuⅡ等酶切后,在琼脂糖凝胶平板与聚丙烯酰胺凝胶柱上进行电泳分析,分别以BamHⅠ与EcoRⅠ双重酶解的λDNA、HindⅢ酶解的T5DNA、HaeⅢ酶解的M13 DNA与HaeⅢ酶解的pBR322 DNA作为泳动的标记,计算出质粒酶解片段的分子量,用此方法算出各片段分子量的总和为15.93×10~6道尔顿,此即为所求的pBR322::Tn233(CH)分子量,将此值减去pBR322的分子置2.87×10~6道尔顿,得到Tn233(CH)的分子量为13.06×10~6道尔顿。电泳结果还表明在Tn233(CH)DNA分子上,BamHⅠ、EcoRⅠ、PstⅠ、HindⅢ与PvuⅡ分别有5、9、1、6、2个切点数。     

大尺蠖核多角体病毒DNA的限制性消化和物理图谱

用5种限制性核酸内切酶消化大尺蠖核多角体病毒(BsNPV)基因组DNA,所得片段数分别是:BamH Ⅰ,9;Bgl Ⅱ,7;Xho Ⅰ,10;HindⅢ 13;EcoR Ⅰ,12,从各个片段的累加测出BsNPV基因组的平均大小约为91,75kb;分子量约为59.60×10~6d。在单酶消化的基础上,用BamH Ⅰ/Bgl Ⅱ双酶消化得到16个片段(即9+7);大小为91,27kb,用、Bgl Ⅱ/Xho Ⅰ双酶消化产生7+10=17个片段,大小为90.04kb,由此组建了这三个酶在BsNPV基因组上的物理图谱。     

棉铃虫单核衣壳核多角体病毒解螺旋酶基因的序列分析

对棉铃虫单核衣壳核多角体病毒（Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus,HaSNPV）基因组中EcoR I-N片段进行序列分析,获得了完整的解螺旋酶基因（hel）,其开放阅读框大小为3762bp,编码一个分子量为146kD的蛋白质。在hel起始密码子ATG上游50位有强晚期启动子转录起始信号ATAAG,在－112位和－189位存在两个TATA box,但未发现早期转录信号CAGT。其在终止密码子下游第12位有一PolyA终止信号AATAAA。在其它真核或原解螺旋酶中存在的7个保守基元（Ⅰ、Ⅰa、Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ）,只有5个（Ⅰ、Ⅰa、Ⅱ、Ⅲ、Ⅳ）在杆状病毒中保守。同源性比较发现,HaSNPV解螺旋酶的氨基酸序列与甜菜夜蛾核多角体病毒（Spodoptera exigus MNPV,SeMNPV）的解螺旋酶具有最高的同源性（66％）,与Xestia c-nigrum颗粒体病毒（XcGV）解螺旋酶的同源性最低（43％）。HaSNPV解螺旋酶基因是第一个报道的单粒包埋核多角体病毒的解螺旋酶基因。     

犬Ⅰ型腺病毒DNA的酶切分析及分子克隆

犬Ⅰ型腺病毒(CAV-1)弱毒用限制性内切酶EcoR Ⅰ,BamH Ⅰ,Pst Ⅰ,Sph Ⅰ和Hind Ⅲ消化分析后其图谱与强毒株相比没有差异。将弱毒DNA用Pst Ⅰ完全消化后以鸟枪法克隆到载体质粒pBluescrip'SK中,经用光生物素标记的CAV-1 DNA杂交筛选以及Pst Ⅰ分析重组质粒证明已将分子量为5.5,3.5,2.85,1.2,0.32和0.28Kb的CAV-1DNA片段克隆到质粒中。克隆到的这些片段将可考虑进一步研究作为探针检测犬及狐狸等野生动物的腺病毒感染。     

大熊猫线粒体DNA的九种限制酶图谱

本文用9种限制性内切酶（BamHⅠ,BglⅠ,BglⅡ,EcoRⅠ,EcoRⅤ,PstⅠ,PvuⅡ,SalⅠ,XhoⅠ）分析大熊猫的线粒体DNA（mtDNA）。构建其中5种酶（BamHⅠ,EcoRⅠ,EcoRⅤ,PstⅠ,PvuⅡ）的mtDNA物理图谱。大熊猫mtDNA的分子大小约为16.4 Kb,酶切位点是随机分布。我们的结果为进一步研究大熊猫mtDNA进化提供了基础资料。     

蜀柏毒蛾核型多角体病毒结构多肽及基因组酶切分析

对蜀柏毒蛾核型多角体病毒(Parocneria orienta Nuclear polyhedrovirus,简称PaorNPV)形态结构、结构多肽、限制性内切酶图谱等特性进行了研究.采用不连续系统垂直板SDS-PAGE分析了PaorNPV的多角体蛋白、病毒粒子结构多肽.应用5种限制性内切酶对PaorNPV基因组DNA进行了酶切分析.结果表明:经热处理的多角体蛋白仅有一条带,分子量为31.5 kD,不经热处理的多角体蛋白有三条带,分子量分别为31.5 kD、29.1 kD、28.6 kD;病毒粒子包含有25种结构多肽,分子量范围在17.6-114.6 kD之间.PaorNPV DNA经BamH I.EcoR I、HindⅢ、Pst I和Xho I酶切分别产生9、12、12、12和14条片段.基因组大小平均为124.6 kb.     

Investigation of the mechanism of temperature sensitivity of the electroreceptors of ampullae of lorenzini

In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980.     

Principles of organization and evolution of systems of regulation of functions

Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms. The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning of physiological systems and organs of the living organism