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1.
Summary DNA sequence analysis of genetic deletions in bacteriophage T7 has shown that these chromosomal rearrangements frequently
occur between directly repeated DNA sequences. To study this type of spontaneous deletion in more quantitative detail synthetic
fragments of DNA, made by hybridizing two complementary oligonucleotides, were introduced into the non-essential T7 gene 1.3
which codes for T7 DNA ligase. This insert blocked synthesis of functional ligase and made the phage that carried an insert
unable to form plaques on a host strain deficient in bacterial ligase. The sequence of the insert was designed so that after
it is put into the T7 genome the insert is bracketed by direct repeats. Perfect deletion of the insert between the directly
repeated sequences results in a wild-type phage. It was found that these deletion events are highly sensitive to the length
of the direct repeats at their ends. In the case of 5 bp direct repeats excision from the genome occurred at a frequency of
less than 10−10, while this value for an almost identical insert bracketed by 10 bp direct repeats was approximately 10−6. The deletion events were independent of a hostrecA mutation. 相似文献
2.
Carboxyl-terminal deletion analysis of the Streptococcus mutans glucosyltransferase-I enzyme 总被引:9,自引:0,他引:9
Sequential deletion of the carboxyl-terminal amino acids (including the six direct repeating units) of the glucosyltransferase-I (GTF-I) enzyme of Streptococcus mutans revealed differential effects on sucrase and GTF activities. Removal of all but one repeating unit resulted in a truncated enzyme with significant sucrase activity but no detectable GTF activity. These results are compatible with the presence of two functional domains in the enzyme. 相似文献
3.
4.
The size distribution of insertions and deletions in human and rodent pseudogenes suggests the logarithmic gap penalty for sequence alignment 总被引:20,自引:0,他引:20
The size distributions of deletions, insertions, and indels (i.e., insertions or deletions) were studied, using 78 human processed pseudogenes and other published data sets. The following results were obtained: (1) Deletions occur more frequently than do insertions in sequence evolution; none of the pseudogenes studied shows significantly more insertions than deletions. (2) Empirically, the size distributions of deletions, insertions, and indels can be described well by a power law, i.e., f
k
= Ck
–b
, where f
k
is the frequency of deletion, insertion, or indel with gap length k, b is the power parameter, and C is the normalization factor. (3) The estimates of b for deletions and insertions from the same data set are approximately equal to each other, indicating that the size distributions for deletions and insertions are approximately identical. (4) The variation in the estimates of b among various data sets is small, indicating that the effect of local structure exists but only plays a secondary role in the size distribution of deletions and insertions. (5) The linear gap penalty, which is most commonly used in sequence alignment, is not supported by our analysis; rather, the power law for the size distribution of indels suggests that an appropriate gap penalty is w
k
= a + b ln k, where a is the gap creation cost and blnk is the gap extension cost. (6) The higher frequency of deletion over insertion suggests that the gap creation cost of insertion (a
i
) should be larger than that of deletion (a
d
); that is, a
i
– a
d
= In R, where R is the frequency ratio of deletions to insertions.
Correspondence to: W.-H. Li 相似文献
5.
In large-scale bioprocesses microbes are exposed to heterogeneous substrate availability reducing the overall process performance. A series of deletion strains was constructed from E. coli MG1655 aiming for a robust phenotype in heterogeneous fermentations with transient starvation. Deletion targets were hand-picked based on a list of genes derived from previous large-scale simulation runs. Each gene deletion was conducted on the premise of strict neutrality towards growth parameters in glucose minimal medium. The final strain of the series, named E. coli RM214, was cultivated continuously in an STR-PFR (stirred tank reactor – plug flow reactor) scale-down reactor. The scale-down reactor system simulated repeated passages through a glucose starvation zone. When exposed to nutrient gradients, E. coli RM214 had a significantly lower maintenance coefficient than E. coli MG1655 (Δms = 0.038 gGlucose/gCDW/h, p < 0.05). In an exemplary protein production scenario E. coli RM214 remained significantly more productive than E. coli MG1655 reaching 44% higher eGFP yield after 28 h of STR-PFR cultivation. This study developed E. coli RM214 as a robust chassis strain and demonstrated the feasibility of engineering microbial hosts for large-scale applications. 相似文献
6.
Courtney L. Jones Teena Bhatla Roy Blum Jinhua Wang Steven W. Paugh Xin Wen Wallace Bourgeois Danielle S. Bitterman Elizabeth A. Raetz Debra J. Morrison David T. Teachey William E. Evans Michael J. Garabedian William L. Carroll 《The Journal of biological chemistry》2014,289(30):20502-20515
Although great advances have been made in the treatment of pediatric acute lymphoblastic leukemia, up to one of five patients will relapse, and their prognosis thereafter is dismal. We have previously identified recurrent deletions in TBL1XR1, which encodes for an F-box like protein responsible for regulating the nuclear hormone repressor complex stability. Here we model TBL1XR1 deletions in B-precursor ALL cell lines and show that TBL1XR1 knockdown results in reduced glucocorticoid receptor recruitment to glucocorticoid responsive genes and ultimately decreased glucocorticoid signaling caused by increased levels of nuclear hormone repressor 1 and HDAC3. Reduction in glucocorticoid signaling in TBL1XR1-depleted lines resulted in resistance to glucocorticoid agonists, but not to other chemotherapeutic agents. Importantly, we show that treatment with the HDAC inhibitor SAHA restores sensitivity to prednisolone in TBL1XR1-depleted cells. Altogether, our data indicate that loss of TBL1XR1 is a novel driver of glucocorticoid resistance in ALL and that epigenetic therapy may have future application in restoring drug sensitivity at relapse. 相似文献
7.
Molecular mapping around the centromere of tomato chromosome 6 using irradiation-induced deletions 总被引:3,自引:0,他引:3
T. B. Liharska J. Hontelez Ab van Kammen Pim Zabel M. Koornneef 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(5-6):969-974
Irradiation-induced deletion mapping was exploited to construct a detailed locus-order map around the centromere of tomato
chromosome 6 (CEN 6). An F1 hybrid heterozygous for the marker loci thiamineless (tl), yellow virescent (yv) and potato leaf (c), and homozygous recessive for the nematode resistance gene mi, was pollinated with γ-irradiated pollen from cultivar VFNT Cherry carrying the wild-type alleles at the corresponding loci.
A dose of 100 Gy was found optimal for inducing mutants. By screening for pseudo-dominant plants showing the marker phenotypes
and/or nematode susceptibility, 30 deletions encompassing one or more of the four loci were detected in the M1 generation. Molecular-marker analysis revealed that 29 of these mutants included the tl and mi loci on the short arm and originated from terminal deletions of different sizes. Remarkably, the breakpoints of these deletions
were not randomly distributed along the short arm but located within the centromeric heterochromatin. Only one yv interstitial deletion and no c mutations on the long arm of the chromosome were detected. Mapping of the various chromosomal breakpoints in the isolated
mutants permitted the resolution of a cluster of molecular markers from the centromeric heterochromatin that was hitherto
unresolvable by genetic linkage analysis. The usefulness of such a deletion-mapping approach for whole-genome mapping is discussed.
Received: 4 March 1997 / Accepted: 2 June 1997 相似文献
8.
Summary The relative rates of point nucleotide substitution and accumulation of gap events (deletions and insertions) were calculated for 22 human and 30 rodent processed pseudogenes. Deletion events not only outnumbered insertions (the ratio being 71 and 31 for human and rodent pseudogenes, respectively), but also the total length of deletions was greater than that of insertions. Compared with their functional homologs, human processed pseudogenes were found to be shorter by about 1.2%, and rodent pseudogenes by about 2.3%. DNA loss from processed pseudogenes through deletion is estimated to be at least seven times faster in rodents than in humans. In comparison with the rate of point substitutions, the abridgment of pseudogenes during evolutionary times is a slow process that probably does not retard the rate of growth of the genome due to the proliferation of processed pseudogenes. 相似文献
9.
Absence of direct repeats flanking transposons resulting from intramolecular transposition 总被引:2,自引:0,他引:2
Tn2660 is an ampicillin-resistance-conferring transposon with a high degree of homology for the transposon Tn3. The nucleotide sequences flanking the termini of Tn2660 have been determined on plasmids inferred to have resulted from both inter- and intramolecular transposition of Tn2660. In all cases, transposition of Tn2660, as of Tn3, creates 5-bp flanking direct repeats, except following intramolecular transposition resulting from trans ligation. In this case, in R6K replicons, the nucleotide sequence between the two Tn2660 elements is stably inverted from the normal orientation, and 5-bp direct repeats do not flank each transposon, but instead flank opposite ends of the two transposon copies. 相似文献
10.