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151.
Takehiro Masumura Daisuke Shibata Takashi Hibino Tomohiko Kato Koichi Kawabe Go Takeba Kunisuke Tanaka Shoji Fujii 《Plant molecular biology》1989,12(2):123-130
Using a rice maturing seed pUC9 expression library, we isolated a cDNA clone corresponding to 10 kDa sulfurrich prolamin by immunoscreening. A longer cDNA clone was obtained from a gtll library by plaque hybridization using this 32P-labeled cDNA as a probe. A polypeptide sequence composed of 134 amino acids was deduced from the nucleotide sequence. A 24 amino acid signal peptide was assigned by computer calculation for the membrane spanning region and Edman sequencing of the purified mature polypeptide. Remarkably, 20% of methionine and 10% of cysteine were found in the mature polypeptide as well as high contents of glutamine, and hydrophobic amino acids. Part of the amino acid sequence was homologous with a conserved cysteine-rich region found in other plant prolamins. Two repeats of amino acid sequence were found in the polypeptide. 相似文献
152.
DNA replication in maize leaf protoplasts 总被引:1,自引:0,他引:1
H. Wang Adrian J. Cutler M. Saleem Larry C. Fowke 《Plant Cell, Tissue and Organ Culture》1989,18(1):33-46
Maize leaf protoplasts were investigated for their metabolic competence and capacity to synthesize DNA. When protoplasts were incubated at elevated temperatures, they exhibited a heat shock response with specific proteins being preferentially synthesized. This indicated that the protoplasts were fully metabolically functional and capable of responding to environmental stimuli. Significant DNA synthesis was observed in these protoplasts after incorporation of 3H-thymidine into chromatin by trichloroacetic acid precipitation and by incorporation of 5-bromo-2-deoxyuridine (BrdU), an analog of thymidine, detected by immunofluorescence. The immunocytochemical method revealed that about 50% of nuclei in the maize leaf protoplasts were labelled after 3 days of culture and that most of these nuclei were labelled as intensely as normal mitotic cells. Aphidicolin, an inhibitor of DNA polymerase-, decreased the percentage of labelled nuclei, demonstrating that the labelling was substantially due to replicative DNA synthesis. However, chromosome condensation was not observed. It is proposed that these protoplasts are capable of DNA synthesis, but incapable of nuclear division. Effects of media additives on the number of nuclei entering S phase in these protoplasts were also assessed by the immunocytochemical method. Inclusion of 80mM Ca2+ in the enzyme solution increased protoplast yield and also appeared beneficial to DNA synthesis. The antioxidant, n-propyl gallate, which was used to stabilize the protoplasts, delayed the onset of DNA synthesis. Arginine and spermidine produced a slight increase in DNA synthesis.Abbreviations BrdU
5-bromo-2-deoxyuridine
- DMSO
dimethyl sulfoxide
- n-PG
n-propyl gallate
- PBS
phosphate-buffered saline
Dedicated to Dr. Friedrich Constabel on the occasion of his 60th birthday 相似文献
153.
Green cybrids with a new nucleus-chloroplast combination cannot be selected after protoplast fusion in the intersubfamilial Nicotiana-Solanum combination. As an approach to overcome the supposed plastomegenome incompatibility, a partial plastome transfer by genetic recombination has been considered. After fusions of protoplasts of a light-sensitive Nicotiana tabacum (tobacco) plastome mutant and lethally irradiated protoplasts of wild-type Solanum tuberosum (potato), a single green colony was recovered among 2.5×104 colonies. The regenerated plants had tobacco-like (although abnormal) morphology, but were normally green, and sensitive to tentoxin, demonstrating chloroplast markers of the potato parent. Restriction enzyme analysis of the chloroplast DNA (cpDNA) revealed recombinant, nonparental patterns. A comparison with physical maps of the parental cpDNA demonstrated the presence of a considerable part of the potato plastome flanked by tobacco-specific regions. This potacco plastome proved to be stable in backcross and backfusion experiments, and normally functional in the presence solely of N. tabacum nucleus. 相似文献
154.
Nora Goosen Harold P. A. Horsman René G. M. Huinen Arjan de Groot Pieter van de Putte 《Antonie van Leeuwenhoek》1989,56(1):85-91
From a gene bank of theAcinetobacter calcoaceticus genome a plasmid was isolated that complements four different classes of PQQ- mutants. Subclones of this plasmid revealed that the four corresponding PQQ genes are located on a fragment of 5 kilobases. The nucleotide sequence of this 5 kb fragment was determined and by means of Tn5 insertion mutants the reading frames of the PQQ genes could be identified. Three of the PQQ genes code for proteins of Mr 29700 (gene I), Mr 10800 (gene II) and Mr 43600 (gene III) respectively. In the DNA region where gene IV was mapped however the largest possible reading frame encodes for a polypeptide of only 24 amino acids. A possible role for this small polypeptide will be discussed. Finally we show that expression of the four PQQ genes inAcinetobacter lwoffi andEscherichia coli lead to the synthesis of the coenzyme in these organisms. 相似文献
155.
Mitochondrial DNA polymorphism in native Philippine cattle based on restriction endonuclease cleavage patterns 总被引:4,自引:0,他引:4
Tomomasa Watanabe Joseph S. Masangkay Shigeharu Wakana Naruya Saitou Takeshi Tomita 《Biochemical genetics》1989,27(7-8):431-438
An analysis of patterns of cleavage of mtDNA by restriction endonucleases was performed for nine individuals from the Philippine
population of native cattle. MtDNA polymorphisms were detected in the restriction patterns generated by the following six
enzymes,BamHI,BglII,EcoRV,HindIII,PstI, andScaI. The restriction patterns showing polymorphisms were distributed nonrandomly among the nine individuals examined from the
Philippine population of native cattle, indicating the existence of two separate types of mtDNA. These two types of mtDNA
are very different from each other, at the level of subspecies. Since the native Philippine cattle are considered to represent
an admixture of European and Indian cattle, the two types of mtDNA must be derived from the mtDNAs of both varieties. The
polymorphic sites in mtDNA have been located on a restriction map, and the nucleotide substitutions at some of the sites have
also been estimated. 相似文献
156.
Methods are presented to extract and purify mitochondrial DNA from the rotifer Brachionus plicatilis. The mtDNA obtained is of sufficient purity for digestion with restriction endonucleases. EcoR I restriction patterns are presented for 4 geographically separated clones. A restriction map based on digestion with 5 different restriction enzymes is included for one of these clones. Finally, use of mtDNA analysis for studies on the population structure and biogeography of rotifers is discussed. 相似文献
157.
158.
Elvira Maniccia-Bozzo Myrna Bueno Espiritu Gurmit Singh 《Molecular and cellular biochemistry》1990,94(1):83-88
The objective of this study was to determine if the nephrotoxic effects induced by cisplatin were correlated to mitochondrial DNA damage. Comparisons were made with the liver since hepatotoxicity is rarely observed. Cisplatin doses of 10, 20 and 40 mg/kg were administered intraperitoneally to C57BL/6J mice. Mitochrondrial DNA was isolated from both the hepatic and renal tissues and quantitated by hybridization with a specific mitochondrial probe. Cisplatin caused differential effects on mouse hepatic and renal mitochondrial DNA. The 10 and 20 mg/kg dose caused an elevation in mitochondrial DNA levels in the hepatic, but no increase in the renal tissue was observed. This is the first study demonstrating an organ specific effect of cisplatin at the DNA level. 相似文献
159.
Elissavet Kardami 《Molecular and cellular biochemistry》1990,92(2):129-135
Summary We have examined the effect of crude cardiac tissue extracts as well as that of several growth factors and triiodothyronin (T3) on DNA synthesis of cardiac myocytes in culture. Extracts from embryonic and adult cardiac tissue stimulated DNA synthesis of myocytes. Atrial myocytes exhibited overall higher degree of stimulation than their ventricular counterparts and extracts from adult atrial tissue had the highest apparent mitogenic activity for atrial myocytes. We have shown that adult heart contains basic fibroblast growth factor (bFGF), especially in the atria [1]. Transforming growth factor (TGF) and insulin-like growth factors (IGFs) are also accumulated in cardiac tissues [2, 3]. We found that bFGF and the IGFs stimulate myocyte cell proliferation and DNA synthesis. These factors also stimulate cardiac non-muscle proliferation, especially in the presence of serum. TGF inhibited proliferation and DNA synthesis and cancelled the effect of bFGF or IGFs on the myocytes. T3 also diminished the bFGF-induced mitogenic stimulation of cardiomyocytes. Our data suggest that these factors may be involved in the regulation of cardiomyocyte proliferation in vivo.Abbreviations bFGF
basic Fibroblast Growth Factor
- BSA
Bovine Serum Albumin
- DM
Defined Medium
- Fes
Fetal calf serum
- FITC
Fluorescein
- IGF
Insulin-like Growth Factor
- IgG
Immunoglobulin
- LI
Labeling Index
- PBS
Phosphate Buffered Saline
- T3
Triiodothyronine
- TGF
Transforming Growth Factor 相似文献
160.
Josée Harel Linda Duplessis Jeffrey S. Kahn Michael S. DuBow 《Archives of microbiology》1990,154(1):67-72
The 37,000 bp double-stranded DNA genome of bacteriophage Mu behaves as a plaque-forming transposable element of Escherichia coli. We have defined the cis-acting DNA sequences required in vivo for transposition and packaging of the viral genome by monitoring the transposition and maturation of Mu DNA-containing pSC101 and pBR322 plasmids with an induced helper Mu prophage to provide the trans-acting functions. We found that nucleotides 1 to 54 of the Mu left end define an essential domain for transposition, and that sequences between nucleotides 126 and 203, and between 203 and 1,699, define two auxiliary domains that stimulate transposition in vivo. At the right extremity, the essential sequences for transposition require not more than the first 62 base pairs (bp), although the presence of sequences between 63 and 117 bp from the right end increases the transposition frequency about 15-fold in our system. Finally, we have delineated the pac recognition site for DNA maturation to nucleotides 32 to 54 of the Mu left end which reside inside of the first transposase binding site (L1) located between nucleotides 1–30. Thus, the transposase binding site and packaging domains of bacteriophage Mu DNA can be separated into two well-defined regions which do not appear to overlap.Abbreviations
attL
attachment site left
-
attR
attachment site right
- bp
base pairs
- Kb
kilobase pair
- nt
nucleotide
- Pu
Purine
- Py
pyrimidine
- Tn
transposable element
State University of New York, Downstate Medical Center, Brooklyn, NY 11204 USA 相似文献