麦迪霉素产生菌具有启动功能的DNA片段的克隆和分析

利用启动子探针质粒载体pIJ486从麦迪霉素产生菌总DNA中克隆得到了一段具有启动功能的DNA片段.通过限制性酶酶切分析,测定插入DNA片段大小为2.3kb.又利用载体pIJ486和pIJ487的新霉素抗性结构基因上游有多酶切点方向相反的性质,分析了插入片段在两个不同方向上的启动能力.结果表明,在两个方向上均有启动功能,但强弱相差六倍.其中在XbaI-HindIII方向上具有较强的启动能力,在变铅青链霉菌中新霉素抗性水平可达20mg/ml以上.进一步对插入片段的三个BamHI小片段进行分析的结果表明,较强启动子区域集中在BamHI-BamHI 0.79kb DNA片段上.     

Nicardipine inhibits axon conduction but causes dual changes of acetylcholine release in the mouse motor nerve

The effects of nicardipine, a dihydropyridine Ca2(+)-channel antagonist, on neuromuscular transmission and impulse-evoked release of acetylcholine were compared with those of nifedipine. In the isolated mouse phrenic nerve diaphragm, nicardipine (50 microM), but not nifedipine (100 microM), induced neuromuscular block, fade of tetanic contraction, and dropout or all-or-none block of end-plate potentials. Nicardipine had no significant effect on the resting membrane potential and the amplitude of miniature end-plate potentials but increased the frequency and caused the appearance of large size miniature potentials. The quantal contents of evoked end-plate potentials were increased. In the presence of tubocurarine, however, nicardipine depressed the amplitude of end-plate potentials. The compound nerve action potential was also decreased. It is concluded that nicardipine blocks neuromuscular transmission by acting on Na+ channels and inhibits axonal conduction. Nicardipine appeared to affect the evoked release of acetylcholine by dual mechanisms, i.e., an enhancement presumably by an agonist action on Ca2+ channels, like Bay K 8644 and nifedipine, and inhibition by an effect on Na+ channels, like verapamil and diltiazem. In contrast with its inactivity on the amplitude of miniature end-plate potentials, depolarization of the end plate in response to succinylcholine was greatly depressed. The contractile response of baby chick biventer cervicis muscle to exogenous acetylcholine was noncompetitively antagonized by nicardipine (10 microM), but was unaffected by nifedipine (30 microM). These results may implicate that nicardipine blocks the postsynaptic acetylcholine receptor channel by enhancing receptor desensitization or by a use-dependent effect.     

Larval development of Crangon hakodatei Rathbun (Decapoda: Crangonidae) reared in the laboratory

Complete larval development of Crangon hakodatei Rathbun isdescribed, based on material hatched in the laboratory fromovigerous females. The species has six zoeal stages and onepostlarval stage. The morphological characters of the larvaland postlarval stages are described with illustrative figuresand compared with those of two congeneric species. The zoealstages of C. hakodatei can be distinguished from those of otherCrangon species in the number of segments of the antennule peduncle,the number of setae on the antennal scale and basis of the maxillipeds,and the stages of appearance of pereiopods. The first zoealstage in the seven species of Crangon are compared and an annotatedkey for distinguishing them is also provided.     

Bacteria Associated with Cysts of the Soybean Cyst Nematode (Heterodera glycines)

The soybean cyst nematode (SCN), Heterodera glycines, causes economically significant damage to soybeans (Glycine max) in many parts of the world. The cysts of this nematode can remain quiescent in soils for many years as a reservoir of infection for future crops. To investigate bacterial communities associated with SCN cysts, cysts were obtained from eight SCN-infested farms in southern Ontario, Canada, and analyzed by culture-dependent and -independent means. Confocal laser scanning microscopy observations of cyst contents revealed a microbial flora located on the cyst exterior, within a polymer plug region and within the cyst. Microscopic counts using 5-(4,6-dichlorotriazine-2-yl)aminofluorescein staining and in situ hybridization (EUB 338) indicated that the cysts contained (2.6 ± 0.5) × 10⁵ bacteria (mean ± standard deviation) with various cellular morphologies. Filamentous fungi were also observed. Live-dead staining indicated that the majority of cyst bacteria were viable. The probe Nile red also bound to the interior polymer, indicating that it is lipid rich in nature. Bacterial community profiles determined by denaturing gradient gel electrophoresis analysis were simple in composition. Bands shared by all eight samples included the actinobacterium genera Actinomadura and Streptomyces. A collection of 290 bacteria were obtained by plating macerated surface-sterilized cysts onto nutrient broth yeast extract agar or on actinomycete medium. These were clustered into groups of siblings by repetitive extragenic palindromic PCR fingerprinting, and representative isolates were tentatively identified on the basis of 16S rRNA gene sequence. Thirty phylotypes were detected, with the collection dominated by Lysobacter and Variovorax spp. This study has revealed the cysts of this important plant pathogen to be rich in a variety of bacteria, some of which could presumably play a role in the ecology of SCN or have potential as biocontrol agents.     

Cigarette smoke-induced bronchoconstriction: causative agents and role of thromboxane receptors

Hong, Ju-Lun, and Lu-Yuan Lee. Cigarette smoke-inducedbronchoconstriction: causative agents and role of thromboxane receptors. J. Appl. Physiol. 81(5):2053-2059, 1996.Inhalation of cigarette smoke induces a biphasicbronchoconstriction in guinea pigs: the first phase is induced by acombination of cholinergic reflex and tachykinins, whereas the secondphase involves cyclooxygenase metabolites (J.-L. Hong, I. W. Rodger,and L.-Y. Lee. J. Appl. Physiol. 78:2260-2266, 1995). This study was carried out to further determinethe causative agents in the smoke and the types of prostanoid receptorsand endogenous prostanoids mediating the bronchoconstriction. Inhalation of 10 ml of high-nicotine cigarette smoke consistently elicited the biphasic bronchoconstriction in anesthetized and artificially ventilated guinea pigs. Pretreatment with hexamethonium (10 mg/kg iv) significantly reduced the first-phase bronchoconstriction but did not have any measurable effect on the second-phase response. Insharp contrast, gas-phase smoke did not elicit any bronchoconstrictive effect. Furthermore, when the animals were challenged with low-nicotine cigarette smoke, only a single second-phase response was evoked, accompanied by increases in thromboxane (Tx)B₂ (a stable metabolite ofTxA₂), prostaglandin (PG)D₂,PGF₂ in the bronchoalveolar lavage fluid. The bronchoconstrictive response induced by low-nicotine smoke was completely prevented by pretreatment with SQ-29548 (0.3 mg/kgiv), a TxA₂-receptor antagonist.These results indicate that 1)nicotine is the primary causative agent responsible for the first-phasebronchoconstriction and 2)nonnicotine smoke particulates evoke the release ofTxA₂,PGD₂, andPGF₂, which act onTxA₂ receptors on airway smoothmuscles and induce the second-phase response to cigarette smoke.

     

Structure and expression of a rice hsp70 gene

A genomic hsp70 gene was isolated from a rice IR36 genomic library and 4 794 bp of the gene have been sequenoed. The 5' flanking region of the gene contained a putative TATA box and a typical heat shock element sequence 5'-CTcgGAAccTTCgAG-3'. The amino acid sequence of the rice HSP70 deduced from the coding region shared 84%-92% homologies with those of HSP70s from other plant species. An intron 1939bp long was identified in the coding region at the codon specifying amino acid 72 (Asp), the similar position introns occurring in other intron-containing hsp70 genes. In addition, another intron of 57 bp was found in the 3'-untranslated region in the rice hsp70 gene. Southern blot hybridization showed that rice hsp70 gene family contained at least three members. Analysis of the RNA leveis with the gene-specific and non-specific probes revealed that the rice hsp70 gene expressed at normal temperature and the expression was enhanced by heat shock treatment.     

梅花鹿甲烷能代谢规律的研究

本文应用ＫＢ－１型呼吸测热装置,结合消化、代谢试验,对梅花鹿（Ｃｅｒｖｕｓｎｉｐｐｏｎ）甲烷能代谢规律进行了研究。结果表明,梅花鹿甲烷能的产生量随其采食量的增加而增加;也随着果食后时间的推移而减少,而且减少的幅度又随采食量的增加而下降;甲烷能的产生量分别占总能食入量、消化能食入量和体增热的６．６１％、８．８３％和１０．８８％;甲烷能的产生量随着日粮蛋白质水平的提高而降低,日粮蛋白质水平每提高１个百分点,甲烷能产生量就降低５８．５８ｋＪ／ｄ;分别以总能食入量（ＧＥＩ）和干物质食入量（ＤＭＩ）为自变量所建立的甲烷能（ＣＨ４Ｅ）估计分别为：ＣＨ４Ｅ（ｋＪ／ｄ）＝０．０７ＣＥＪ（ｋＪ／ｄ）－１０１．０４（ｎ＝１２,ｒ＝０．９４４,Ｐ＜０．０１）ＣＨ４Ｅ（ｋＪ／ｄ）＝９８．７８＋１．０５ＤＭＩ（ｇ／ｄ）（ｎ＝１２,ｒ＝０．９４２,Ｐ＜０．０１）     

视黄醇结合蛋白的结构与功能

视黄醇结合蛋白（RBP）是视黄醇转运的载体蛋白,作为结合小分子流水物质的载体蛋白家族（lipocalin）的一个重要成员,其结构与功能的研究正受到国外学者的重视,文章介绍了视黄醇结合蛋白的性质、结构研究进展,讨论了视黄醇结合蛋白与前蛋白和受体相互作用的位点和结构特点．