苏云金芽胞杆菌鲇泽亚种HD—133 cry1D的表达调控

利用PCR扩增基因crylD启动子及上游区片段,在测序的基础上构建含crylD—lacZ融合基因穿梭质粒,导入不同遗传背景的苏云金芽胞杆菌菌株中,并以crylAb—lacZ融合基因为对照测定β—半乳糖苷酶活性,检测启动子上游区的作用。结果表明,crylD—lacZ和crylAb—lacZ融合基因在不同遗传背景的菌株中表达完全不同,也许一些宿主专一性的因子参与了转录调控;而在同一菌株中CerylD-lacZ和cryl Ab-lacZ的表达差异是由于上游区的不同以及竞争有限的σ因子所致。利用PCR定点诱变技术突变其SD序列GGGGA为GGAGG后,CerylD-lacZ融合基因的表达提高了1．0—1．6倍。表明GGAGG是苏云金芽胞杆菌合适的SD序列,也揭示了不合适的SD序列是crylD表达量低的原因之一。     

A new role for BiP: closing the aqueous translocon pore during protein integration into the ER membrane.

In mammalian cells, most membrane proteins are inserted cotranslationally into the ER membrane at sites termed translocons. Although each translocon forms an aqueous pore, the permeability barrier of the membrane is maintained during integration, even when the otherwise tight ribosome-translocon seal is opened to allow the cytoplasmic domain of a nascent protein to enter the cytosol. To identify the mechanism by which membrane integrity is preserved, nascent chain exposure to each side of the membrane was determined at different stages of integration by collisional quenching of a fluorescent probe in the nascent chain. Comparing integration intermediates prepared with intact, empty, or BiP-loaded microsomes revealed that the lumenal end of the translocon pore is closed by BiP in an ATP-dependent process before the opening of the cytoplasmic ribosome-translocon seal during integration. This BiP function is distinct from its previously identified role in closing ribosome-free, empty translocons because of the presence of the ribosome at the translocon and the nascent membrane protein that extends through the translocon pore and into the lumen during integration. Therefore, BiP is a key component in a sophisticated mechanism that selectively closes the lumenal end of some, but not all, translocons occupied by a nascent chain. By using collisional quenchers of different sizes, the large internal diameter of the ribosome-bound aqueous translocon pore was found to contract when BiP was required to seal the pore during integration. Therefore, closure of the pore involves substantial conformational changes in the translocon that are coupled to a complex sequence of structural rearrangements on both sides of the ER membrane involving the ribosome and BiP.     

The carboxyl-terminal isoforms of smooth muscle myosin heavy chain determine thick filament assembly properties.

The alternatively spliced SM1 and SM2 smooth muscle myosin heavy chains differ at their respective carboxyl termini by 43 versus 9 unique amino acids. To determine whether these tailpieces affect filament assembly, SM1 and SM2 myosins, the rod region of these myosin isoforms, and a rod with no tailpiece (tailless), were expressed in Sf 9 cells. Paracrystals formed from SM1 and SM2 rod fragments showed different modes of molecular packing, indicating that the tailpieces can influence filament structure. The SM2 rod was less able to assemble into stable filaments than either SM1 or the tailless rods. Expressed full-length SM1 and SM2 myosins showed solubility differences comparable to the rods, establishing the validity of the latter as a model for filament assembly. Formation of homodimers of SM1 and SM2 rods was favored over the heterodimer in cells coinfected with both viruses, compared with mixtures of the two heavy chains renatured in vitro. These results demonstrate for the first time that the smooth muscle myosin tailpieces differentially affect filament assembly, and suggest that homogeneous thick filaments containing SM1 or SM2 myosin could serve distinct functions within smooth muscle cells.     

A New Marine Reptile (Sauropterygia) from New Zealand: Further Evidence for A Late Cretaceous Austral Radiation of Cryptoclidid Plesiosaurs

Kaiwhekea katiki gen. et sp. nov. represents the first described cryptoclidid plesiosaurian from New Zealand. It is one of the largest cryptoclidids known, at a length of over 6.5 m, and represents the third reported genus of austral Late Cretaceous cryptoclidids. Kaiwhekea katiki is from siltstones of the Katiki Formation, upper Haumurian Stage (Cenomanian–Maastrichtian; c. 69–70 Ma) of coastal Otago, South Island, New Zealand. In the Late Cretaceous, the locality lay close to the polar circle. The holotype and only known specimen is an articulated skeleton with skull, preserved mostly as natural molds, but which lacks the forelimbs and pectoral girdle. The skull is relatively large and possesses several distinct characters, including a substantial, deep, jugal. There are about 43 upper and 42 lower teeth in each jaw quadrant; all are homodont, slim, and slightly recurved, lacking prominent ornament. Kaiwhekea probably took single soft-bodied prey. Based on cranial structure, it clearly belongs with the Cryptoclididae, but is not certainly close to the southern Late Cretaceous cryptoclidids Morturneria (Seymour Island, Antarctica) and Aristonectes (Chile, Argentina).     

Distribution and cycling of C, N, Ca, Mg, K and P in three pristine, old-growth forests in the Cordillera de Piuchué, Chile

We assessed a number of biomass and soil parameters in order to examinerelationships among nutrient availability, forest productivity and vegetationpatterns in two old-growth forested watersheds in a pristine montane landscapeon Isla de Chiloé, Chile. We selected watersheds in both gymnosperm- andangiosperm-dominated forests and determined tree species, d.b.h. and health forall trees < 2 cm d.b.h. in plots established at 50m intervals. Soils were sampled at two depths in each plot andanalyzed for total C and N, and for exchangeable Ca, K, Mg andresin-extractableP. Allometric relationships and vegetation nutrient concentrations were used todetermine above-ground pools from the vegetation survey data. Growth rates werederived from increment core measures. Soil pools of most elements measuredappear adequate to support forest growth indefinitely. Mineralized nitrogen,which is similar in quantity to the annual demand for nitrogen from the soil isthe exception, consistent with the possibility of N limitation in two of theforest types studied. A third type, an evergreen broadleaved forest, appears torequire substantially more nitrogen than would appear to be available from netmineralization measurements. Productivity per unit of nitrogen required fromthesoil is quite high, largely as a consequence of the evergreen habit of thespecies in these forests. Compared to other temperate montane forests in theNorthern Hemisphere, nutrient pools and cycling characteristics were found tobemostly similar across forest types, in spite of considerable variation invegetation and soils.     

Chemolithoautotrophic Nitrifiers in the Phyllosphere of a Spruce Ecosystem Receiving High Atmospheric Nitrogen Input

Evidence is presented for the first time that chemolithoautotrophic ammonia oxidizers (CAO) and chemolithoautotrophic nitrite oxidizers (CNO) colonize in appreciable cell numbers the phyllosphere of spruce trees in a forest ecosystem exposed for decades to high levels of atmospheric nitrogen (The Höglwald Forest, Bavaria, Germany). The results strongly indicate that both, CAO and CNO are predominantely located inside the spruce needles, most likely within the stomatal cavity. These results are further supported by field experiments of NH₃ uptake into twigs on intact spruce trees in the presence and absence of 10 Pa acetylene, an inhibitor of the ammonia monooxygenase of CAO. It is clearly demonstrated for the first time that in situ uptake of NH₃ from the atmosphere into spruce needles exposed to high levels of atmospheric N is not catalyzed exclusively by the tree, but is the result of combined activities of both, the spruce trees and the chemolithoautotrophic nitrifiers colonizing the needles.     

The Californian urban butterfly fauna is dependent on alien plants

Abstract. Using the unusually well-documented butterfly fauna of Davis, Yolo County, California, it is shown that the mainly native species commonly observed in gardens breed mostly or entirely on alien plants, especially naturalized weeds. Over 40% of the fauna has no known native hosts in the urban–suburban environment. Were certain alien weeds to be eradicated or their abundance greatly reduced, the urban-suburban butterfly fauna would disappear. This might be regarded as an unfortunate, and perhaps intolerable, side-effect of such programs.     

Pollination and phenology of flowers in the canopy of two contrasting rain forest types in Amazonia,Colombia

The main objective of this investigation was to study the pollination characteristics of two types of Amazonian rain forest at plant community level. Seasonally inundated forest was compared with upland (tierra firme) forest. The study focused on plant species in the canopy. The pollination spectra show that in both forests most canopy trees and lianas are pollinated by small bees, large bees, butterflies or by small, relatively unspecialized insects. In the upland forest small bees are the most important pollinators (32% of all species of trees and lianas are pollinated by them), whereas large bees are predominant in the floodplain (22%). Other pollinators, like hummingbirds, bats, moths, and beetles are less common (>10%), but always somewhat more important in the flood plain than in the upland forest. Bees are the most common pollinators of epiphytes. In the flood plain forest, flies are also important as epiphyte pollinators (19%), whereas in the upland forest hummingbirds pollinate more epiphytes. The phenological patterns are quite similar in both the upland and the flood plain. We found a peak in flowering in the transition period between the wet and the dry season. Flowering activity was lowest during the wet season. Differentiation in sexual systems was correlated with life form. Dioecy and monoecy were found mostly among tree species. Most species of all life forms though were hermaphroditic. No difference with respect to the relative importance of sexual systems was found between the two forest types.     

Differences in Composition and Mucosal Adhesion of Bifidobacteria Isolated from Healthy Adults and Healthy Seniors

Fifty-one Bifidobacterium strains were isolated from the feces of healthy adults (30–40 years old) and seniors (older than 70 years of age). B. adolescentis, B. breve, B. infantis, and B. longum were isolated from the healthy adults and B. adolescentis and B. longum from elderly subjects. The tested bacteria bound, in vitro, to intestinal mucus in a strain dependent manner. The strains isolated from healthy adults, and especially B. adolescentis, bound better to intestinal mucus than those isolated from seniors. These results indicate that the mucosal adhesive properties of the human Bifidobacterium flora were reduced with the aging of the host. This shift to a Bifidobacterium flora with reduced adhesive abilities may explain the decrease in bifidobacteria levels in the intestinal microflora of aging people. Received: 7 February 2001 / Accepted: 3 April 2001