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61.
62.
Cloning and Mapping of a Novel Nodulation Region from Bradyrhizobium japonicum by Genetic Complementation of a Deletion Mutant 总被引:4,自引:2,他引:2 下载免费PDF全文
The phenotypes of a set of Bradyrhizobium japonicum 110 mutants with large deletions in the region of symbiotic gene cluster I were tested. The majority of the mutants showed a delayed nodulation on soybean and, by mixed-infection experiments, were found to be strongly reduced in their competitiveness. Phenotypic comparison of mutants with different deletion endpoints allowed a preliminary localization of two genomic regions, called nod-1 and nod-2, which were required for normal nodulation on soybean. Loss of nod-1 was found to result in a Nod− phenotype on cowpea, mung bean, and siratro. A recombinant cosmid was identified which fully restored nodulation ability of a mutant lacking nod-1. Using Tn5-containing derivatives and subclones of this cosmid for complementation, we delimited the nod-1 region to a DNA segment of 3.1 to 3.5 kilobase pairs. 相似文献
63.
A Balland T Faure D Carvallo P Cordier P Ulrich B Fournet H de la Salle J P Lecocq 《European journal of biochemistry》1988,172(3):565-572
A stable transformed cell line constitutively expressing human factor IX has been established. Wild-type Chinese hamster ovary cells (CHO cells) were transformed using a polycistronic expression vector carrying a previously isolated factor IX cDNA and a selection gene encoding the Escherichia coli xanthine-guanine phosphoribosyl transferase. One clone, CHO 622.4, contains a high number of genomically integrated plasmids and secretes 1-3 mg factor IX l-1 day-1 into the culture medium with a biological activity ranging from 25% to 40%. The recombinant molecule was purified either by conventional chromatography or by immunoaffinity chromatography using antibodies specific to a calcium-induced factor IX conformer. The purified recombinant protein migrates as a single band with the same mobility as that of natural factor IX on SDS/polyacrylamide gels. N-terminal sequencing shows tow differently processed forms of recombinant factor IX: whereas the majority of the zymogen is correctly processed, approximately 20% of the purified recombinant molecule contains an 18-amino-acid NH2-extension corresponding to the precursor form of factor IX. Analysis of the 4-carboxyglutamic acid content indicates a high but incomplete carboxylation (70%) of the recombinant molecule as compared to natural factor IX. The carbohydrate composition of both the natural and recombinant molecules has been determined. Both molecules have a N-glycan structure of similar complexity, indicating that factor IX contains all the information to direct the same glycosylation pattern in human liver cells and in an unrelated cell line such as CHO-K1. 相似文献
64.
Expression of the chemically synthesized gene for ribonuclease T1 in Escherichia coli using a secretion cloning vector 总被引:3,自引:0,他引:3
R Quaas Y McKeown P Stanssens R Frank H Bl?cker U Hahn 《European journal of biochemistry》1988,173(3):617-622
The gene for ribonuclease T1 from Aspergillus oryzae has been chemically synthesized using the segmental support technique. An Escherichia coli clone producing the ribonuclease at high levels was constructed by linking the gene downstream to the region coding for the signal peptide of the OmpA protein (a major outer membrane protein of E. coli), using the secretion cloning vector pIN-III-ompA2. This strategy was employed in order to circumvent a possible toxic effect of the gene product on the host cell. Active ribonuclease containing four additional amino acids at the N-terminus could be isolated from the periplasmic fraction of the host. The final yield after purification was 20 mg enzyme/l liquid culture. With respect to immunological, catalytic and specific behaviour, no qualitative differences could be detected between the enzyme from the over-producing E. coli strain and ribonuclease T1 isolated from A. oryzae. 相似文献
65.
Peter Berthold und Ulrich Querner 《Journal of Ornithology》1988,129(3):372-375
Summary In 9 hand-raised Garden Warblers, Zugunruhe was continuously recorded in registration cages with movable perches on microswitches in a constant 12:12 hr light-dark cycle throughout the first fall migratory period. In addition, video-records were taken during 23 nights under infrared illumination involving more than 150 hours. The main results are: (1) infrared illumination did not affect Zugunruhe or other events and thus is a useful method, (2) over 90 % of the Zugunruhe recorded with video is wing whirring (i. e. intense wing beating in a sitting position), the rest is hopping and flying, (3) an interval of the customarily used half-an-hour-with-Zugunruhe equals 14.1 min of wing whirring, (4) 35.9 % of the Zugunruhe recorded as wing whirring is missed in a registration cage with one movable perch. This loss is reduced to about 5 % if two movable perches are used, (5) the experimental birds produced in the average 352.1 ± 170.51 (SD) hours with Zugunruhe which equals about 165 hrs of wing whirring. When this value is multiplied by the average flight speed of the species during migration (about 25–30 km/h) a theroretical flight distance of about 4800 km is obtained. This is just the distance from the south German breeding grounds via Spain to the estuary of the river Niger which is situated within the center of the population-specific winter quarters. Zugunruhe in that species thus appears with some restrictions to be wandering by wing whirring in a sitting position. 相似文献
66.
67.
68.
Ulrich Melcher In Seong Choe Genevieve Lebeurier Ken Richards Richard C. Essenberg 《Molecular & general genetics : MGG》1986,203(2):230-236
Summary Some cauliflower mosaic virus (CaMV) alleles are selectively lost during growth of the virus in mixedly infected turnip plants. Viral DNA from plants co-inoculated with DNA of the cabbage S isolate and infectious cabbage S DNA with an extra EcoRI restriciion site lacked the extra site. The EcoRI allele was also lost in most plants co-inoculated with a non-infectious mutant of cabbage S DNA while little selective allele loss was observed with two other non-infectious mutant DNAs. Plants co-inoculated with DNAs of closely-related isolates (CM4-184 and W) contained both parental viral DNAs and some DNAs with characteristics of both parents. Interference, scored as a reduced frequency of infection or a delay in symptom appearance relative to plants inoculated with wild-type DNA, occurred when plants were inoculated with wild-type and mutant DNAs covalently attached to one another in partial dimer plasmid DNAs. Similarities in the conditions leading to selective allele loss and those leading to interference suggest that both may have been due to active gene conversion between CaMV DNA molecules. 相似文献
69.
M P Kirpichnikov A V Kurochkin B K Chernov F Buck K D Hahn 《Molekuliarnaia biologiia》1986,20(6):1562-1569
An oligodeoxyribonucleotide composed of 17 residues, d(TATCACCGCCAGAGGTA), and a complementary chain were synthesized. Their duplex was identical with the operator OR1, the binding site for bacteriophage lambda cro and c1 repressors. The 1H NMR spectra (500 MHz) of the duplex imino and aromatic protons were studied at 10, 20 and 25 degrees C. Signals from the imino protons of complementary base pairs and from the C2 protons of adenine (with the exception of the duplex terminal nucleotides) were assigned using the NOE technique and the known characteristics of short DNA fragment melting. No signals from the imino protons of the terminal base pairs were detected even at 10 degrees C due to fraying which increased as the temperature was raised. The assignment of signals can be used to identify centers of interaction between the operator OR1 and repressors, as well as to study possible local changes in DNA geometry. 相似文献
70.
Philippe Szankasi Christof Gysler Ulrich Zehntner Urs Leupold Jürg Kohli Peter Munz 《Molecular & general genetics : MGG》1986,202(3):394-402
Summary Recombination between dispersed yet related serine tRNA genes of Schizosaccharomyces pombe does occur during mitosis but it is approximately three orders of magnitude less frequent than in meiosis. Two mitotic events have been studied in detail. In the first, a sequence of at least 18 nucleotides has been transferred from the donor sup3 gene on the right arm of chromosome I to the related acceptor gene sup12 on the left arm of the same chromosome, thereby leading to the simultaneous change of 8 bp in the acceptor gene. This event must be explained in terms of recombination rather than mutation. It is assumed that it represents mitotic gene conversion, although it was not possible to demonstrate that the donor gene had emerged unchanged from the event. The second case reflects an interaction between sup9 on chromosome III and sup3 on chromosome I. Genetic and physical analysis allows this event to be described as mitotic gene conversion associated with crossingover. The result of this event is a reciprocal translocation. No further chromosomal aberrations were found among an additional 700 potential intergenic convertants tested. Thus intergenic conversion is much less frequently associated with crossingover than allelic conversion. However, the rare intergenic conversion events associated with crossingover provide a molecular mechanism for chromosomal rearrangements. 相似文献