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21.
Molecular cloning and expression of the xylanase gene of alkalophilic Bacillus sp. strain C-125 in Escherichia coli. 总被引:6,自引:1,他引:5
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The gene for xylanase A of alkalophilic Bacillus sp. strain C-125 was cloned in Escherichia coli with pBR322. The plasmid pCX311 contained 2.6- and 2.0-kilobase-pair HindIII fragments. The characteristics of the purified pCX311-encoded xylanase were the same as those of purified xylanase A from alkalophilic Bacillus sp. strain C-125. 相似文献
22.
23.
Molecular cloning and expression of cellulase genes of alkalophilic Bacillus sp. strain N-4 in Escherichia coli. 总被引:14,自引:8,他引:6
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The genes for cellulases of alkalophilic Bacillus sp. strain N-4 were cloned in Escherichia coli with pBR322. Plasmids pNK1 and pNK2 were isolated from the transformants producing carboxymethyl cellulase, and the carboxymethyl cellulase genes cloned were in 2.0- and 2.8-kilobase-pair HindIII fragments, respectively. On the DNA level, the pNK1 fragment had a different restriction map from that of the pNK2 fragment, but the genomic hybridization experiments showed partial homology among these fragments. A total of 74 and 34% of the enzyme activities were observed in the periplasmic space of E. coli carrying the plasmids pNK1 and pNK2 , respectively. The carboxymethyl cellulase thus produced had broad pH activity curves (pH of 5 to 10.9) and was stable up to 75 degrees C. 相似文献
24.
Shin-ichi Miyoshi Katsumi Sugiyama Hiroshi Furuta Noriko Miyoshi Sumio Shinoda 《FEMS microbiology letters》1986,34(3):301-304
Abstract Vibrio vulnificus protease (VVP) stimulated histamine release from isolated mast cells in a dose- and temperature-dependent manner within a range of 0.2–4.0 μ g/0.5 ml. Histamine release was accompanied by degranulation, and no leakage of lactate dehydrogenase from cells was observed, indicating that the histamine release was not due to cytolysis but to exocytosis. This release, completed within 30 s at 37°C, suggested that the mechanism of action of VVP on mast cells is different from that of other proteases, such as trypsin or α-chymotrypsin, which release histamine from the cells slowly. 相似文献
25.
26.
Excretion of the penicillinase of an alkalophilic Bacillus sp. through the Escherichia coli outer membrane. 总被引:7,自引:3,他引:4
Two plasmids containing the penicillinase gene of alkalophilic Bacillus sp. strain 170, pEAP1 and pEAP2, were constructed. Most of the penicillinase produced by Escherichia coli, which carried these plasmids, was found in the culture medium. This excretion is caused by the cloned DNA fragment which contains some component that changes the outer membrane of E. coli. 相似文献
27.
Selective Inhibition of Reovirus Ribonucleic Acid Synthesis by Cycloheximide 总被引:27,自引:18,他引:9
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Cycloheximide, at a concentration of 10 mug/ml, rapidly blocked protein synthesis in L cells infected with reovirus. When the drug was added before 5 hr postinfection, synthesis of both single- and double-stranded varieties of virus-specific ribonucleic acid (RNA), which normally commences between 6 and 7 hr after infection, was blocked. When the cycloheximide was added at 9 hr after infection, uptake of uridine-H(3) into RNA, for the succeeding 6 hr at least, was similar to that of an infected culture without the drug. This latter uptake of uridine-H(3) in the presence of cycloheximide was largely into single-stranded RNA, since double-stranded RNA synthesis was rapidly and markedly inhibited by the cycloheximide. Single-stranded RNA formed in the presence of cycloheximide was found not to be a precursor of viral progeny, double-stranded RNA. Synthesis of double-stranded RNA in the infected cell probably requires prior synthesis of a new protein, which has a rapid rate of turnover. The possibility that formation of single-stranded RNA is preceded by synthesis of a second new protein is discussed. 相似文献
28.
T Kaneko K B Song T Hamamoto T Kudo K Horikoshi 《Journal of general microbiology》1989,135(12):3447-3457
The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from the alkalophilic Bacillus sp. strain no. 17-1 was cloned in Escherichia coli. The cloned CGTase gene consisted of a single open reading frame which would encode a polypeptide of 713 amino acids, and the first 27 amino acid residues comprised a signal peptide. The nucleotide sequence and the amino acid sequence of this CGTase (CGTase 17-1) gene had strong homology with those of the CGTase (CGTase 38-2) gene previously cloned in our laboratory from the alkalophilic Bacillus sp. strain no. 38-2, although the enzymic properties of the CGTase 17-1 were distinct from those of the CGTase 38-2. To analyse those enzymic properties further, we constructed 12 chimeric CGTases using three restriction nuclease sites and compared the enzymic properties of the chimeric CGTases. The N-terminal part of the enzyme was important for heat stability, and the pH-activity profile was influenced by both the N- and the C-terminal parts. A third segment was less important for enzymic properties. 相似文献
29.
Copulatory behavior unrelated to conception is sometimes observed in some non-human primates including the Japanese monkey.
In the present study, the authors examined whether a mature follicle or a newly formed fresh corpus luteum was observed in
the ovaries of female Japanese monkeys which displayed the copulatory behavior unrelated to conception.
Post-conception copulatory behaviors were observed in three out of four females usually kept in individual cages in an air-conditioned
room, and in two out of three females without infants kept in an outdoor group cage. However, neither a mature Graafian follicle
nor a fresh corpus luteum formed newly after conception was observed in any of these females by laparoscopic examinations
conducted immediately after termination of the copulatory behavior. In females with infants born in the preceding birth season,
copulatory behaviors were observed in three out of four females kept in the outdoor group cage, and in two out of four females
in a free-ranging troop. Ovulation was confirmed in one case out of the three kept in the outdoor group cage, but neither
a mature follicle nor a newly formed corpus luteum was observed in the remaining four females.
These findings suggest that copulatory behavior in the Japanese monkey is not always controlled by the development of a follicle
or ovulation in the ovary. 相似文献
30.
Two dynorphin-degrading cysteine proteases, I and II, were extracted with Triton X-100 from neuroblastoma cell membrane, isolated from accompanying dynorphin-degrading trypsin-like enzyme by affinity chromatography on columns of soybean trypsin inhibitor-immobilized Sepharose and p-mercuribenzoate-Sepharose, and separated by ion-exchange chromatography on diethylaminoethyl (DEAE)-cellulose and TSK gel DEAE-5PW columns. Cysteine protease II was purified further by hydroxyapatite chromatography and gel filtration. The molecular weights of cysteine proteases I and II were estimated to be 100,000 and 70,000, respectively, by gel filtration. Both of the enzymes, were inhibited by p-chloromercuribenzoate, N-ethylmaleimide, and high-molecular-weight kininogen, but not or only slightly inhibited by diisopropylphosphorofluoridate, antipain, leupeptin, E-64, calpain inhibitor, and phosphoramidon. Cysteine protease I cleaved dynorphin(1-17) at the Arg6-Arg7 bond with the optimum pH of 8.0, whereas II cleaved dynorphin(1-17) at the Lys11-Leu12 bond and the Leu12-Lys13 bond with the optimum pH values of 8.0 and 6.0, respectively. These bonds corresponded to those that had been proposed as the initial sites of degradation by neuroblastoma cell membrane. Cysteine protease I was further found to show strict specificity toward the Arg-Arg doublet, when susceptibilities of various peptides containing paired basic residues were examined as substrates for the enzyme. 相似文献