江浙蝮蛇蛇毒蛋白C激活因子基因的克隆及测序

从江浙蝮蛇毒腺中抽提总RNA，RT－PCR进行体外扩增，获得江浙蝮蛇蛇毒蛋白C激活因子基因，克隆至pGEX-5X-3载体中，对3个重组克隆分别作DNA全序列分析，通过遗传密码推导出相应的氨基酸序列，与其它已知的丝氨酸复白酶蛇毒蛋白的氨基酸作比较，其中许多上氨基酸有很强的同源性，该基因的成功克隆，不仅推导出江浙蝮蛇蛇毒蛋白C激活因子的蛋白质序列，也为进一步开展江浙蝮蛇蛇毒蛋白C激活因子蛋白质工程的研究工作打下良好的基础。     

Effects of glucose,glutamine, and malate on the metabolism of spodoptera frugiperda clone 9 (sf9) cells

Optimal design and operation of bioreactors for insect cell culture is facilitated by functional relations providing quantitative information on cellular metabolite consumption kinetics, as well as on the specific cell growth rates (μ_G). Initial specific consumption rates of glucose, malate, and oxygen, and associated changes in μ_G, were measured forSpodoptera frugiperda clone 9 (Sf9) cells grown in batch suspension culture in medium containing 7–35 mM glucose, 0–16 mM malate, and 4–16 mM glutamine. The initial specific glucose consumption rate (q _G ) could be described by a modified Michaelis-Menten equation treating malate as a “competitive” inhibitorK ₁ = 6.5 mM) and glutamine as a “noncompetitive” inhibitorK _I = 14 mM) ofq _G , with aK _m of 7.1 mM for glucose. All three carbon sources were found to increase μ_G in a saturable manner, and a modified Monod equation was employed to describe this relationship (μ_Gmax = 0.047 h^-1). The initial specific oxygen consumption rate (q_O2) in Sf9 cells could be related to μ_G by the maintenance energy model, and it was calculated that, under typical culture conditions, about 15–20% of the cellular energy demand comes from functions not related to growth. Fitted parameters in mathematical expression for μg: K₄, Monod constant for glucose (mM); K₅, modified Monod constant for malate (mM); K₆, Monod constant for glutamine (mM); m_o2, specific consumption rate of oxygen by the cells under zero-growth conditions (nmol/cell/h); q_F, initial specific fumarate production rate (nmol/cell/ h);q _G , initial specific glucose consumption rate (nmol/cell/h); q_Gmax, maximum initial specific glucose consumption rate (nmol/cell/h);q _M , initial specific malate consumption rate (nmol/cell/h); q_o2, initial specific oxygen consumption rate (nmol/cell/h); Y_o2, cell yield on oxygen (cells/nmol); μ, initial specific cell growth rate (h^-1); μ_g, initial specific cell growth rate (h^-1); μG_max, maximum initial specific cell growth rate (h^-1).     

Metrizability and Coconnectedness

Solving the problem stated in Sichler and Trnková, Topol. Its Appl., 142: 159–179, 2004, we construct metrics μ, ν on a set P such that the spaces X=(P,μ) and Y=(P,ν) have the same monoid of all continuous selfmaps, the space Y is coconnected (in the sense that every continuous map Y×Y→Y depends on at most one coordinate) while X is not. Also, properties of the forgetful functors Metr → Unif → Top are investigated for the “simultaneous variant” of the above problem. Supported by the Grant Agency of Czech Republic under grant 201/06/0664 and by the project of Ministry of Education of Czech Republic MSM 0021620839.     

小菜蛾颗粒体病毒DNA限制性酶切分析及基因组DNA片段克隆

小菜蛾颗粒体病毒ＤＮＡ用ＢａｍＨＩ和ＸｈｏＩ酶切，经０．７５％琼脂糖凝胶电泳，分别得到１２条带和８条带，平均分子量为１１３．０ｋｂ．用Ｓｕｐｅｒｃｏｓ１Ｃｏｓｍｉｄ作载体，将Ｓａｕ３ＡＩ部分消化的ＰｘＧＶ－ＤＮＡ随机片段插入该载体的ＢａｍＨＩ酶切位点，转化于ＸＬ１－Ｂｌｕｅ受体菌，经Ａｍｐ平板筛选转化子，挑出２５６个Ａｍｐ＋菌落，以３２Ｐ－ｄＣＴＰ标记的ＰｘＧＶ－ＤＮＡ为探针，斑点杂交筛选得到５９个阳性重组体，再经电泳检测，得到了５９个不同大小的ＰｘＧＶ－ＤＮＡ片段克隆．     

On the Length of Scales of Computability Potentials for n-Element Algebras

We find the length of scales of computability potentials for n-element algebras. As a consequence, we find the length of the filter generated by the clone of a discriminant function in the lattice of clones of functions on an n-element set.     

B组轮状病毒WH-1株nsp2基因的原核表达与抗体制备

根据B组轮状病毒(GBRV)WH-1株nsp2基因的全序列,设计引物,用PCR的方法扩增得到nsp2基因的编码区.将其片段克隆到原核表达载体pGEX-KG上,经IPTG诱导,在E.coliDH5α菌株中得到高效表达.表达的GST融合蛋白大小为61×103,经SDS-PAGE分离纯化,免疫小白鼠制备了多克隆抗体.抗体经1∶3 000倍稀释后用于Western Blot分析,获得特异性显色信号.所表达的蛋白和制备的抗体可用于蛋白结构和功能的进一步研究.     

The clone of operations preserving a cycle with loops

We consider all the binary relations on a k-element set which, when viewed as directed graphs, consist of a k-cycle and some loops. If k ≥ 3 and the relation has at least two loops, we show that it is only preserved by essentially unary operations. In all other cases, the relation is preserved by operations that depend on a greater number of variables. Received March 9, 2006; accepted in final form September 14, 2007.     

The join of two minimal clones and the meet of two maximal clones

We construct two minimal clones on any finite set such that the join of the two clones contains all operations. Dually, we exhibit two maximal clones on any finite set with at least three elements such that the intersection of the two clones is the trivial clone containing projections only. Received October 20, 1998; accepted in final form December 16, 1998.     

Generating primitive positive clones

Suppose is a set of operations on a finite set A. Define PPC() to be the smallest primitive positive clone on A containing . For any finite algebra A, let PPC#(A) be the smallest number n for which PPC(CloA) = PPC(Clo _n A). S. Burris and R. Willard [2] conjectured that PPC#(A) ≤|A| when CloA is a primitive positive clone and |A| > 2. In this paper, we look at how large PPC#(A) can be when special conditions are placed on the finite algebra A. We show that PPC#(A) ≤|A| holds when the variety generated by A is congruence distributive, Abelian, or decidable. We also show that PPC#(A) ≤|A| + 2 if A generates a congruence permutable variety and every subalgebra of A is the product of a congruence neutral algebra and an Abelian algebra. Furthermore, we give an example in which PPC#(A) ≥|A| - 1)² so that these results are not vacuous. Received August 30, 1999; accepted in final form April 4, 2000.