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61.
Ole Olsen Rainer Borriss Ortwin Simon Karl Kristian Thomsen 《Molecular & general genetics : MGG》1991,225(2):177-185
Summary Hybrid (1-3,1-4)--glucanase genes were constructed by extension of overlapping segments of the (1-3,1-4)--glucanase genes from Bacillus amyloliquefaciens and B. macerans generated by the polymerase chain reaction (PCR). Four hybrid genes were expressed in Escherichia coli cells. The mature hybrid enzymes contain a 16, 36, 78, or 152 amino acid N-terminal sequence derived from B. amyloliquefaciens (1-3,1-4)--glucanase followed by a C-terminal segment derived from B. macerans (1-3,1-4)--glucanase. Biochemical characterization of parental and hybrid enzymes shows a significant increase in thermostability of three of the hybrid enzymes when exposed to an acidic environment thus combining two important enzyme characteristics within the same molecule. At pH 4.1, 85%-95% of the initial activity was retained after 1 h at 65° C in contrast to 5% and 0% for the parental enzymes from B. amyloliquefaciens and B. macerans. After 60 min incubation at 70° C, pH 6.0, the parental enzymes retained 5% or less of the initial activity whilst one of the hybrids still exhibited 90% of the initial activity. Of the parental enzymes B. macerans (1-3,1-4)--glucanase had the lower specific activity while the hybrid enzymes exhibited specific activities that were 1.5- to 3-fold higher. These experimental results demonstrate that exchange of homologous gene segments from different species may be a useful technique for obtaining new and improved versions of biologically active proteins.Abbreviations AMY
mature form of Bacillus amyloliquefaciens (1-3,1-4)--glucanase;
- MAC
mature form of B. macerans (1-3,1-4)--glucanase
- SUB
mature form of B. subtilis (1-3,1-4)--glucanase
- H(A16-M), H(A36-M), H(A78-M), H(A107-M), H(A152-M)
mature forms of hybrid enzymes having 16, 36, 78, 107, 152 N-terminal amino acids, respectively, derived from AMY with the remaining amino acids derived from MAC 相似文献
62.
Summary Freshwater snails and anuran tadpoles have been suggested to have their highest population densities in ponds of intermediate size where abiotic disturbance (e.g. desiccation) is low and large predators absent. Both snails and tadpoles feed on periphytic algae and, thus, there should be a large potential for competitive interactions to occur between these two distantly related taxa. In a field experiment we examined the relative strength of competition between two closely related snail species, Lymnaea stagnalis and L. peregra, and between L. stagnalis and tadpoles of the common frog, Rana temporaria. Snail growth and egg production and tadpole size at and time to metamorphosis were determined. Effects on the common food source, periphyton, were monitored with the aid of artificial substrates. Periphyton dry weight was dramatically reduced in the presence of snails and/or tadpoles. There were no competitive effects on growth or egg production of the two snail species when they were coexisting. Mortality of L. peregra was high (95%) after reproduction, but independent of treatment. Growth of L. stagnalis was reduced only at the highest tadpole densities, whereas egg production was reduced both by intraspecific competition and by competition with tadpoles. Differences in egg production were retained after tadpole metamorphosis. Tadpole larval period increased, weight of metamorphosing frogs decreased and growth rate was reduced as a function of increasing tadpole density. However, contrary to expectation, snails had a positive effect on tadpole larval period, weight and growth rate. Further, in experimental containers without snails there was a dense growth of the filamentous green alga Cladophora sp. We suggest that the facilitative effects of snails on tadpoles are due to an indirect mutualistic mechanism, involving competition between food sources of different quality (microalgae and Cladophora sp.) and tadpoles being competitively dominant over snails for the preferred food source (microalgae). In the presence of tadpoles snails will be forced to feed on low-quality Cladophora, increasing nutrient turnover rates, which results in enhanced productivity of microalgae, increasing tadpole food resources. Thus, tadpoles have a negative effect on snails through resource depression, while snails facilitate tadpole growth through an indirect enhancement of food availability. 相似文献
63.
64.
65.
66.
Spatial restriction of AChR gene expression to subsynaptic nuclei. 总被引:21,自引:0,他引:21
67.
Ten different Polycomb group genes are required for spatial control of the abdA and AbdB homeotic products. 总被引:27,自引:0,他引:27
Mutations in genes of the Polycomb (Pc) group cause abnormal segmental development due to ectopic expression of the homeotic products of the Antennapedia and bithorax complexes. Here the requirements for Pc group genes in controlling the abdA and AbdB products of the bithorax complex are described. Embryos containing mutations in the genes Polycomb (Pc), extra sex combs (esc), Enhancer of zeste [E(z)], polyhomeotic (ph), Sex comb on midleg (Scm), Polycomb-like (Pcl), Sex comb extra (Sce), Additional sex combs (Asx), Posterior sex combs (Psc) and pleiohomeotic (pho) were examined. In every case, both abdA and AbdB are expressed outside of their normal domains along the anterior-posterior (A-P) axis, consistent with these Pc group products acting in a single pathway or molecular complex. The earliest detectable ectopic expression is highest in the parasegments immediately adjacent to the normal expression boundary. Surprisingly, in the most severe Pc group mutants, the earliest ectopic AbdB is distributed in a pair-rule pattern. At all stages, ectopic abdA in the epidermis is highest along the anterior edges of the parasegments, in a pattern that mimics the normal abdA cell-specific pattern. These examples of highly patterned mis-expression show that Pc group mutations do not cause indiscriminate activation of homeotic products. We suggest that the ectopic expression patterns result from factors that normally activate abdA and AbdB only in certain parasegments, but that in Pc group mutants these factors gain access to regulatory DNA in all parasegments. 相似文献
68.
Eric Giraud Alain Brauman Simon Keleke Bertrand Lelong Maurice Raimbault 《Applied microbiology and biotechnology》1991,36(3):379-383
Summary An amylolytic lactic acid bacterium identified as Lactobacillus plantarum was isolated from cassava roots (Manihot esculenta var. Ngansa) during reting. The amylolytic enzyme synthesized was an extracellular -amylase with an optimum pH of 5.0 and an optimum temperature of 55° C. Cultured on starch, the strain displayed a growth rate of 0.43 h–1, a biomass yield of 0.19 g·g–1 and a lactate yield of 0.81 g·g–1. The growth kinetics were similar on starch and glucose. Sufficient enzyme was synthesized and starch hydrolysis was not a limiting factor for growth. Biosynthesis of the enzyme was observed when the glucose concentration was less than 6.7 g·l–1 and reached up to 4 IU·ml–1 at the end of the fermentation.
Offprint requests to: M. Raimbault 相似文献
69.
The sperm nuclei of Aulacomya ater, family Mitylidae, contain three proteins (X, Aa5 and Aa6) which are specific to this cell type coexisting with a set of five somatic-type histones. Information about the chromatin structure resulting from this kind of association is scarce. Therefore, we have probed the structure of this sperm chromatin through digestion with micrococcal nuclease in combination with salt fractionation. The data obtained have allowed us to propose a nucleosomal arrangement for this chromatin. However, two types of nucleosomes would be present in agreement with their protein components. 相似文献
70.
N. Aste C. Viglietti-Panzica A. Fasolo C. Andreone H. Vaudry G. Pelletier G. C. Panzica 《Cell and tissue research》1991,265(2):219-230
Summary In the present study, we have demonstrated, by means of the biotin-avidin method, the widespread distribution of neuropeptide Y (NPY)-immunoreactive structures throughout the whole brain of the Japanese quail (Coturnix coturnix japonica). The prosencephalic region contained the highest concentration of both NPY-containing fibres and perikarya. Immunoreactive fibres were observed throughout, particularly within the paraolfactory lobe, the lateral septum, the nucleus taeniae, the preoptic area, the periventricular hypothalamic regions, the tuberal complex, and the ventrolateral thalamus. NPY-immunoreactive cells were represented by: a) small scattered perikarya in the telencephalic portion (i.e. archistriatal, neostriatal and hyperstriatal regions, hippocampus, piriform cortex); b) medium-sized cell bodies located around the nucleus rotundus, ventrolateral, and lateral anterior thalamic nuclei; c) small clustered cells within the periventricular and medial preoptic nuclei. The brainstem showed a less diffuse innervation, although a dense network of immunopositive fibres was observed within the optic tectum, the periaqueductal region, and the Edinger-Westphal, linearis caudalis and raphes nuclei. Two populations of large NPY-containing perikarya were detected: one located in the isthmic region, the other at the boundaries of the pons with the medulla. The wide distribution of NPY-immunoreactive structures within regions that have been demonstrated to play a role in the control of vegetative, endocrine and sensory activities suggests that, in birds, this neuropeptide is involved in the regulation of several aspects of cerebral functions.Abbreviations
AA
archistriatum anterius
-
AC
nucleus accumbens
-
AM
nucleus anterior medialis
-
APP
avian pancreatic polypeptide
-
CNS
centrai nervous system
-
CO
chiasma opticum
-
CP
commissura posterior
-
CPi
cortex piriformis
-
DIC
differential interferential contrast
-
DLAl
nucleus dorsolateralis anterior thalami, pars lateralis
-
DLAm
nucleus dorsolateralis anterior thalami, pars medialis
-
E
ectostriatum
-
EW
nucleus of Edinger-Westphal
-
FLM
fasciculus longitudinalis medialis
-
GCt
substantia grisea centralis
-
GLv
nucleus geniculatus lateralis, pars ventralis
-
HA
hyperstriatum accessorium
-
Hp
hippocampus
-
HPLC
high performance liquid chromatography
-
HV
hyperstriatum ventrale
-
IF
nucleus infundibularis
-
IO
nucleus isthmo-opticus
-
IP
nucleus interpeduncularis
-
IR
immunoreactive
-
LA
nucleus lateralis anterior thalami
-
LC
nucleus linearis caudalis
-
LFS
lamina frontalis superior
-
LH
lamina hyperstriatica
-
LHRH
luteinizing hormone-releasing hormone
-
LoC
locus coeruleus
-
LPO
lobus paraolfactorius
-
ME
eminentia mediana
-
N
neostriatum
-
NC
neostriatum caudale
-
NPY
neuropeptide Y
-
NIII
nervus oculomotorius
-
NV
nervus trigeminus
-
NVI
nervus facialis
-
NVIIIc
nervus octavus, pars cochlearis
-
nIV
nucleus nervi oculomotorii
-
nIX
nucleus nervi glossopharyngei
-
nBOR
nucleus opticus basalis (ectomamilaris)
-
nCPa
nucleus commissurae pallii
-
nST
nucleus striae terminalis
-
OM
tractus occipitomesencephalicus
-
OS
nucleus olivaris superior
-
PA
palaeostriatum augmentatum
-
PBS
phosphate-buffered saline
-
POA
nucleus praeopticus anterior
-
POM
nucleus praeopticus medialis
-
POP
nucleus praeopticus periventricularis
-
PP
pancreatic polypeptide
-
PYY
polypeptide YY
-
PVN
nucleus paraventricularis magnocellularis
-
PVO
organum paraventriculare
-
R
nucleus raphes
-
ROT
nucleus rotundus
-
RP
nucleus reticularis pontis caudalis
-
Rpc
nucleus reticularis parvocellularis
-
RPgc
nucleus reticularis pontis caudalis, pars gigantocellularis
-
RPO
nucleus reticularis pontis oralis
-
SCd
nucleus subcoeruleus dorsalis
-
SCv
nucleus subcoeruleus ventralis
-
SCNm
nucleus suprachiasmaticus, pars medialis
-
SCNl
nucleus suprachiasmaticus, pars lateralis
-
SL
nucleus septalis lateralis
-
SM
nucleus septalis medialis
-
Ta
nucleus tangentialis
-
TeO
tectum opticum
-
Tn
nucleus taeniae
-
TPc
nucleus tegmenti pedunculo-pontinus, pars compacta
-
TSM
tractus septo-mesencephalicus
-
TV
nueleus tegmenti ventralis
-
VeL
nucleus vestibularis lateralis
-
VLT
nucleus ventrolateralis thalami
-
VMN
nucleus ventromedialis hypothalami
A preliminary report of this study was presented at the 15th Conference of European Comparative Endocrinologists, Leuven, Belgium, September 1990 相似文献