Production of cyclomaltodextrin glucanotransferase of Bacillus circulans var. alkalophilus ATCC21783 in B. subtilis

Summary The cyclomaltodextrin glucanotransferase (CGTase, E.C. 2.4.1.19) gene from an alkalophilic Bacillus circulans var. alkalophilus ATCC21783 was cloned into Escherichia coli and B. subtilis. When cloned from E. coli to B. subtilis, the entire insert containing the CGTase gene was, depending on the plasmid construction, either unstable or the recombinant B. subtilis did not secrete the enzyme in significant amounts. To achieve efficient enzyme production in B. subtilis, the gene was placed under the control of the B. amyloliquefaciens -amylase promoter. In one of the constructions, both the promoter and the signal sequence of the gene were replaced with those of B. amyloliquefaciens, whereas in another construction only the promoter area was exchanged. The recombinant B. subtilis clones transformed with these plasmid constructions secreted CGTase into the culture medium 14 times as much as did the parental strain in shake flask cultures. In fermentor cultures in an industrially feasible medium the enzyme production was substantially higher, yielding 1.2 g/l of CGTase, which is about 33 times the amount of the enzyme produced by the parental strain in corresponding fermentations. Both of the plasmid constructions were stable when grown over 50 generations without antibiotic selection.     

稻田微蛛优势种形成的若干环境因子分析

草间小黑蛛(Erigone graminicolum)和食虫沟瘤蛛(Ummeliata insecticeps)都有较强的耐寒、耐饥能力和较大的捕食量,在-3—0℃时能存活3天左右,平均耐饥历期大于30天,对稻褐飞虱的平均最大捕食量分别为6.3头/天和6.1头/天;Holling模型分别为N_A(草)=0.9886N_0/(1+0.09N_0)和N_A(食)=1.0234N_0/(1+0.101N_0),它们的抗药能力相差无几。但它们对湿度要求不同,前者较后者耐干旱。草间小黑蛛耐干历期雌蛛为27.2天,雄蛛为9.7天;耐湿历期雌蛛为53.2天,雄蛛为46.2天。食虫沟瘤蛛耐干历期雌蛛为7.7天,雄蛛为4.2天;耐湿历期雌蛛为55.4天,雄蛛为49.0天。温度相同湿度不同时它们的竞争能力不同,湿度较小时草间小黑蛛取胜,湿度较大时食虫沟瘤蛛取胜。它们在稻田中成为优势种的条件,主要取决于稻田生境与周围环境的干湿状况。     

狭蚤属一新种记述(蚤目:多毛蚤科)

我们从陕西省南部大巴山(安康地区平利县千家坪林场,位于东经109°15′,北纬31°50′)采获一批蚤类标本,经鉴定其中有狭蚤属 Stenoponia Jordan et Rothschild,1911新种。根据其模式产地,命名为大巴山狭蚤,兹记述如下。 大巴山狭蚤 Stenoponia dabashanensis新种 鉴别特征 新种烦栉刺数和下唇须仅一节,与上海狭蚤 Stenoponia shanghaiensis Liu et Wu 1960 兰狭蚤 S. coclcstis Jordan et Rothschild, 1911 和短距狭蚤S. formozovi Ioff et Tiflov, 1934接近。但下列特征可以区别:1)第9腹板后臂近端部     

Inhibition of receptor-bound urokinase by plasminogen-activator inhibitors

Urokinase-type plasminogen activator (uPA) binds to a specific receptor on various cell types, the bound molecule retaining its enzymatic activity against plasminogen. We have now investigated whether receptor-bound uPA also retains the ability to react with and be inhibited by plasminogen activator inhibitors (PAI-1 and PAI-2). uPA bound to its receptor on human U937 monocyte-like cells was inhibited by PAI-1 (in its active form in the presence of vitronectin fragments) with an association rate constant of 4.5 x 10(6) M-1 s-1, which was 40% lower than that obtained for uPA in solution (7.9 x 10(6) M-1 s-1). The inhibition of uPA by PAI-2 was decreased to a similar extent by receptor binding, falling from 5.3 x 10(5) to 3.3 x 10(5) M-1 s-1. Stimulation of U937 cells with phorbol 12-myristate 13-acetate was accompanied by a further reduction in receptor-bound uPA inhibition by PAI-1 and PAI-2 to 1.7 x 10(6) and 1.1 x 10(5) M-1 s-1, respectively. These constants although lower than those for uPA in solution still represent rather rapid inhibition of the enzyme, and demonstrate that uPA bound to its specific cellular receptor remains available for efficient inhibition by PAI's, which may therefore play a major role in controlling cell-surface plasminogen activation and extracellular proteolytic activity.     

Interactions among coexisting larval Odonata: an in situ experiment using small enclosures

Field experiments using small replicated enclosures focused on interactions between larval populations of Epitheca cynosura and Ladona deplanata (Odonata: Anisoptera) — two species that emerge in early spring. The presence of Epitheca reduced the total biomass of Ladona, but Ladona had no significant effect on Epitheca. These early-emerging species reduced the biomass of small instars of late-emerging Anisoptera which colonized enclosures during the experiments; and the late-emerging Anisoptera seem to have inhibited colonization by Zygoptera larvae. Results are consistent with the importance of predatory (cannibalism or mutual predation) interactions in this community.     

Extraction and immunochemical assays of a tubulin-like factor in cotton seedlings

Antibodies to tubulin were prepared in rabbits by immunization with reduced-carboxymethylated calf-brain tubulin. In immunodiffusion tests the antibodies showed full cross reactivity with the immunogen as well as with native calf-brain tubulin. The same antibodies showed cross reactivity with a factor in extract of cotton (Gossypium hirsutum L.) cotyledons but there was no full immunological identity between calf-brain tubulin and this factor. A solid-phase radioimmunoassay for quantitative estimation of this plant tubulin-like factor was developed. It measured the binding of antibodies to immobilized calf-native tubulin. Competition between the unknown soluble tubulin-like factor, and immobilized tubulin was assayed at serum dilution of 1:50. Extraction conditions which preserved the antigenic properties of the tubulin-like factor from cotton cotyledons were defined. The radioimmunoassay measured quantities of the tubulin-like factor in the range of 0.1–10 g-equivalents of calf-brain tubulin. Immediately after homogenization of the tissue only 25% of the total amount of tubulin-like activity was present in soluble form, while most of it remained in the insoluble fraction. Apparent maximal solubilization was achieved spontaneously 10 h after homogenization or by treatment with guanidine hydrochloride. These results indicate that in this material, tubulin is not released immediately by homogenization but remains assembled in microtubules and-or in a bound or sequestered form.Abbreviations NRS normal rabbit serum - RCM reduced-carboxymethylated     

Chilling Injury in Cotton {Gossypium hirsutum L.): Light Requirement for the Reduction of Injury and for the Protective Effect of Abscisic Acid

Pretreatment by darkness increased chilling (4°C) injuryin whole cotton (Gossypium hirsutum L.) seedlings and isolatedcotyledonary tissue. Addition of sucrose in the dark periodprevented the effect of darkness. Application of the photosyntheticinhibitor DCMU in light simulated the effect of darkness. ABA(10^–5 M) decreased chilling injury when applied in lightas a pretreatment before the onset of chilling. The same pretreatmentin darkness was almost ineffective, unless sucrose was added.ABA applied in light together with DCMU was ineffective in decreasingchilling injury. Lower light intensity resulted in increasedchilling injury and a decreased effect of ABA in the preventionof chilling injury. The antimicrotubular drug colchicine increased the chillinginjury. Pretreatment with ABA in light decreased the chillingand colchicine injury while the same pretreatment in darknesswas ineffective. These results suggest that a deficiency of a photosyntheticproduct increases the chilling sensitivity of the tissue. ABAapparently increases chilling resistance through a metabolicprocess which depends on photosynthetic activity. ³ Incumbent of the Seagram Chair in Plant Sciences (Received November 20, 1980; Accepted January 31, 1981)     

Site-specific integration of an F'' lac pro factor in the region of the replication origin (oriC) of E. coli

Summary An episome, F 128, which carries approximately 8x10⁴ base pairs of chromosomal DNA homologous to the lac pro region of the E. coli chromosome, has been found to integrate into the oriC region of the chromosome in a site specific reaction. While the event appears to be recA-dependent, no homology between the episome and this region of the chromosome was detected. The Hfr strains formed result from the integration of intact F 128 molecules. The structure of the Hfr strains generated has been determined and their transfer properties analyzed.