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71.
V. S. Gupta M. S. Dhar B. G. Patil G. S. Narvekar S. R. Ra Wat P. K. Ranjekar 《Journal of biosciences》1990,15(4):261-269
Rice long repetitive DNA (9–20 kbp) reassociating at Cot 50 M.s was cloned in pBR325. Out of several recombinants (Camr Ampr Tets), only a few were selected randomly for further characterization. The insert size in all these clones was 3–4 kbp. Restriction
enzyme analysis showed the absence ofEcoRI andBclI sites, presence of a singlePstI andPvuII site and multiple sites forAluI in 3 clones namely pRLl, pRL7 and pRL10.
TheBamHI-PstI fragment of about 0.4 kbp in the pRL7 insert DNA (pRL7-0.4 kbp) was subcloned in M13mpl8 and partially sequenced using Sanger’s
dideoxynucleotide chain termination method. Dot matrix comparison of this sequence with rice rDNA sequences revealed low homology
with the 25 S rDNA sequence of rice, however, hybridisation did not indicate any homology. 相似文献
72.
73.
Transforming growth factor alpha and a PC12-derived growth factor induce neurites in PC12 cells and enhance the survival of embryonic brain neurons. 总被引:1,自引:0,他引:1 下载免费PDF全文
We have identified and characterized a 5000-Da protein that induces neurite outgrowth from PC12 pheochromocytoma cells, enhances the survival of embryonic rat brain neurons in primary culture, and induces the multiplication of embryonic rat brain astrocytes in primary culture. The factor is produced by a flat cell PC12 variant that expresses the activated ras oncogene after transfection of the gene. The factor resembles transforming growth factor alpha (TGF alpha) and epidermal growth factor (EGF) in that it induces anchorage-independent colony formation of normal rat kidney cells in soft agar and competes with EGF for binding to the EGF receptor. Rat TGF alpha and human TGF alpha also induce neurite outgrowth from PC12 and enhance the survival of embryonic brain neurons. The PC12 variant-derived factor can be distinguished from TGF alpha and EGF immunologically and by migration rates on reversed-phase high-performance liquid chromatography. 相似文献
74.
野生大豆基因文库的构建 总被引:4,自引:0,他引:4
以氯化铯密度梯度离心法纯化噬菌体λEMBL4,将纯化的EMBL4 DNA用BamH1/SalI双酶切制成载体。用CTAB(十六烷基三甲基溴化铵)法提取野生大豆(种名待定)大分子DNA,Sau3A部分酶解,从琼脂糖凝胶中回收10—22kb“目的”DNA片段,与载体连接,体外包装成重组噬菌体。所得重组子值为8×10(?)pfu(噬菌斑形成单位),达到了构建野生大豆基因文库要求的理论值。以栽培大豆7S贮藏蛋白a′-cDNA作探针,用噬菌斑原位杂交法从文库中筛选出一个阳性克隆。 相似文献
75.
Elizabeth S. Woo Yukihiro Kondo Simon C. Watkins Dale G. Hoyt John S. Lazo 《Experimental cell research》1996,224(2):365
Metallothionein (MT), a major zinc-binding intracellular protein thiol, has been associated with cytoprotection from heavy metals, antineoplastic drugs, mutagens, and cellular oxidants. Despite its small mass (7 kDa), nuclear partitioning of MT has been observed in both normal and malignant tissues. The factors controlling MT sequestration are unknown. Thus, we examined the regulation of MT subcellular distribution in human cancer cell lines that exhibit prominent nuclear MT. The nuclear disposition of MT was unaltered during cell cycle passage in synchronized cells. MT redistributed to the cytoplasm when cells were exposed to reduced temperature. Cytoplasmic redistribution was also seen in DU-145 and HPC36M prostatic cancer cells after ATP depletion, but not in PC3-MA2 and SCC25/CP cells. Pretreatment with 10 μMCdCl2did not significantly alter MT distribution but did render all cells sensitive to cytoplasmic redistribution after either reduced temperature or ATP depletion. Thus, nuclear retention of MT is energy requiring and this ability of MT to accumulate in subcellular compartments against its concentration gradient may be important in the capacity of MT to supply Zn or other metals to target sites within the cell. 相似文献
76.
Carlos Hermenegildo Goizane Marcaida Carmina Montoliu Santiago Grisolía María-Dolores Miñana Vicente Felipo 《Neurochemical research》1996,21(10):1237-1244
We proposed that acute ammonia toxicity is mediated by activation of NMDA receptors. To confirm this hypothesis we have tested
whether different NMDA receptor antagonists, acting on different sites of NMDA receptors, prevent death of mice induced by
injection of 14 mmol/Kg of ammonium acetate, a dose that induces death of 95% of mice. MK-801, phencyclidine and ketamine,
which block the ion channel of NMDA receptors, prevent death of at least 75% of mice. CPP, AP-5, CGS 19755, and CGP 40116,
competitive antagonists acting on the binding site for NMDA, also prevent death of at least 75% of mice. Butanol, ethanol
and methanol which block NMDA receptors, also prevent death of mice. There is an excellent correlation between the EC50 for preventing ammonia-induced death and the IC50 for inhibiting NMDA-induced currents. Acute ammonia toxicity is not prevented by antagonists of kainate/AMPA receptors, of
muscarinic or nicotinic acetylcholine receptors or of GABA receptors. Inhibitors of nitric oxide synthase afford partial protection
against ammonia toxicity while inhibitors of calcineurin, of glutamine synthetase or antioxidants did not prevent ammonia-induced
death of mice. These results strongly support the idea that acute ammonia toxicity is mediated by activation of NMDA receptors. 相似文献
77.
J. Planas P. Ra˚dström F. Tjerneld B. Hahn-Hägerdal 《Applied microbiology and biotechnology》1996,45(6):737-743
In order to enhance the productivity of lactic acid and reduce the end-product inhibition of fermentation, the partitioning
and growth of four different strains of lactic acid bacteria in three different aqueous two-phase systems were studied. Polyethyleneglycol/
dextran, polyethyleneglycol/hydroxypropyl starch polymer (HPS), and a random copolymer of ethylene oxide and propylene oxide
(EO-PO)/HPS were used as polymer systems. One strain each of Lactococcus lactis subsp. lactis and of Lactobacillus delbrueckii subsp. delbrueckii partitioned completely to the interface and bottom phase in two-phase systems with low polymer concentrations of EO-PO/HPS100
and EO-PO/ HPS200. The growth and production of lactic acid by two of three L. lactis strains in a two-phase system with 5.5% (w/w) EO-PO and 12.0% (w/w) HPS100 were reduced by less than 10% compared with a
reference fermentation in a normal growth medium. The viability of L. lactis subsp. lactis ATCC 19435 was maintained for at least 50 h and with four top-phase replacements during extractive fermentation in the EO-PO/HPS100
system. Moreover, when cell density reached the stationary phase in the first extractive fermentation, the lactate production
in this aqueous two-phase system was maintained.
Received: 2 October 1995/Received revision: 16 January 1996/Accepted: 22 January 1996 相似文献
78.
Chae Oh Lim Soo In Lee Woo Sik Chung Sung Han Park Inhwan Hwang Moo Je Cho 《Plant molecular biology》1996,30(2):373-379
A cDNA encoding a new phytocystatin isotype named BCPI-1 was isolated from a cDNA library of Chinese cabbage flower buds. The BCPI-1 clone encodes 199 amino acids resulting in a protein much larger than other known phytocystatins. BCPI-1 has an unusually long C-terminus. A BCPI-1 fusion protein expressed in Escherichia coli strongly inhibits the enzymatic activity of papain, a cysteine proteinase. Genomic Southern blot analysis revealed that the BCPI gene is a member of a small multi-gene family in Chinese cabbage. Northern blot analysis showed that it is differentially expressed in the flower bud, leaf and root. 相似文献
79.
Treatment of the A-ring aromatic steroids estrone 3-methyl ether and β-estradiol 3, 17-dimethyl ether with Mn(CO)5+BF4− in CH2Cl2 yields the corresponding [(steroid)Mn(CO)3]BF4 salts 1 and 2 as mixtures of and β isomers. The X-ray structure of [(estrone 3-methyl ether)Mn(CO)3]BF4 · CH2Cl2 (1) having the Mn(CO)3 moiety on the side of the steroid is reported: space group P21 with a=10.3958(9), b=10.9020(6), c=12.6848(9) Å, β=111.857(6)°, Z=2, V=1334.3(2) Å3, calc=.481 cm−3, R=0.0508, and wR=0.0635. The molecule has the traditional ‘piano stool’ structure with a planar arene ring and linear Mn---C---O linkages. The nucleophiles NaBH4 and LiCH2C(O)CMe3 add to [(β-estradiol 3,17-dimethyl ether)Mn(CO)3]BF4 (2) in high yield to give the corresponding - and β-cyclohexadienyl manganese tricarbonyl complexes (3). The nucleophiles add meta to the arene -OMe substituent and exo to the metal. The and β isomers of 3 were separated by fractional crystallization and the X-ray structure of the β isomer with an exo-CH2C(O)CMe3 substituent is reported (complex 4): space group P212121 with a=7.5154(8), b=15.160(2), c=25.230(3) Å, Z=4, V=2874.4(5) Å3, calc=1.244 g cm−3, R=0.0529 and wR2=0.1176. The molecule 4 has a planar set of dienyl carbon atoms with the saturated C(1) carbon being 0.592 Å out of the plane away from the metal. The results suggest that the manganese-mediated functionalization of aromatic steroids is a viable synthetic procedure with a range of nucleophiles of varying strengths. 相似文献
80.
Summary To isolate a novel gene that contains an SH2 domain, we devised a rapid and nonradioactive cDNA library screening method using polymerase chain reaction (PCR). For PCR amplification, we designed degenerate oligonucleotide primers from the multialigned DNA sequences of SH2 domains. This method offers an inexpensive and efficient approach for the isolation of clones of interest from cDNA libraries. 相似文献