Showing papers in "Gene in 1984"

TL;DR: A method is described for the rapid generation and cloning of deletion derivatives well-suited for the sequencing of long stretches of DNA based on two useful features of exonuclease III: processive digestion at a very uniform rate and failure to initiate digestion at DNA ends with four-base 3'-protrusions.

TL;DR: A new method for in vitro insertional mutagenesis of genes cloned in Escherichia coli that makes use of the omega fragment, a 2.0-kb DNA segment consisting of an antibiotic resistance gene flanked by short inverted repeats carrying transcription and translation termination signals and synthetic polylinkers.

TL;DR: This finding has been exploited to identify the coding sequence of the viomycin phosphotransferase gene of Streptomyces vinaceus, and an easily applied computer program has been written to carry out and display such analyses.

TL;DR: The versatility of insertional inactivation of beta-galactosidase activity for subcloning and sequencing has been enhanced by combining a chemically synthesized oligonucleotide which specifies nine 6-bp-cutter restriction sites to create a set of highly versatile cloning sites.

TL;DR: Several new variants of the tetracycline-resistance transposon Tn10 are described which are more useful than the wild-typetransposon for many types of genetic and physical analysis of bacteria.

TL;DR: In this article, a plasmid cloning vector, pGB2, was constructed from the Escherichia coli pSC101 and showed no sequence homology, as detected by DNA-DNA hybridization, to several widely used vectors such as pBR322, pUC8 and phage lambda L47.1.

TL;DR: The properties of transposon Tn5 that render it useful for in vivo mutagenesis of cloned DNA sequences are reviewed and a methods section is included which outlines precisely how to carry out transposition frequency, insertional specificity, polarity and stability of Tn 5 insertion mutations.

TL;DR: If a DNA fragment containing a promoter is inserted into one of the cloning sites upstream from the antibiotic genes, cells carrying such plasmids acquire resistance to cam or tet, and using these vectors two restriction fragments that contain promoters were identified.

TL;DR: With this method 1 ng of active enzyme can easily be detected and both prokaryotic and eukaryotic cell extracts can be examined, and changes in the size of enzymatically active proteins can be determined.

TL;DR: The beta-galactosidase fusion vector pMC1403 has been modified to include the unique cloning sites EcoRI, SmaI, BamHI, SalI, AccI, PstI and HindIII.

TL;DR: The expression of the cloned Saccharomyces cerevisiae URA3 gene in Escherichia coli on both plasmid and phage vectors was studied and many of the common yeast vector plasmids are now completely known at the level of DNA sequence.

TL;DR: Two new shuttle vectors have been constructed by fusing the Escherichia coli plasmid pUC9 with the Staphylococcus aureus plasmids pU110 and pC194, which replicate in both E. coli and Bacillus subtilis and contain seven restriction sites within a part of the lacZ gene.

TL;DR: The complete nucleotide sequence of a 2.9-kb DNA fragment containing the CDC2 gene-complementing activity from Schizosaccharomyces pombe has been determined and results strongly suggest that the two genes code for similar functions.

TL;DR: A one-step purification method of hybrid proteins exhibiting β-galactosidase activity, based on affinity chromatography in the presence of high salt concentration, is described, which can be used to obtain antibodies against the foreign portion of the protein fusion.

TL;DR: Novel bacterial resistance genes were cloned and expressed as dominant selection markers in mammalian cells and plasmid DNA sequences were identified by Southern blot analysis of high Mr DNA isolated from transformed cells.

TL;DR: A protocol for the rapid restriction mapping of phage lambda clones has been developed and the restriction map can be directly determined from the "ladder" of partial digestion products.

TL;DR: This result confirms the theory that the intergenic region consists of two domains: one domain being a segment involved in phage morphogenesis and the other being a region of functional origin which interferes with M13 replication.

TL;DR: The gene organization of HBV DNA was found to be well conserved irrespective of subtype, and the direct repeat of the undecanucleotide sequence near the 5' ends of the short (S) and long (L) strands ofHBV DNA and the two small direct repeats between both 5' end were found toBe characteristic structures.

TL;DR: By insertion of a DNA fragment, containing the phage lambda pR promoter and the pM-promoted cI857 allele of the lambda repressor gene, in plasmid R1 upstream of the replication control genes, cloning vectors have been constructed which are present in one copy per chromosome at temperatures below 37 degrees C, and which display uncontrolled replication at 42 degrees C.

TL;DR: The promoter region from the cloned glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Saccharomyces cerevisiae has been characterized and demonstrated to contain all sequences necessary for promoter function in vivo.

TL;DR: A rat insulin gene which was fused to Escherichia coli signals for the initiation of translation could not be retained when expressed from the strong rrnB ribosomal RNA promoters or an induced trp/lac (= tac) hybrid promoter, but when the latter promoter was repressed by transformation into a lac-repressor-overproducing strain, the insulin gene fragment could be retained.

TL;DR: A comparison of the promoter region for PGK with other promoters on the X-chromosome reveals homology with the promoter for HPRT, but not with the operator for factor IX.

TL;DR: A detailed restriction enzyme map of M-ABA virus DNA reveals the close similarity to isolates from other sources and stresses the importance of using cloning vectors that can be propagated in recA- Escherichia coli hosts.

TL;DR: Analysis of introns in fungal mitochondrial genes and the self-splicing intron of the large ribosomal RNA of several species of Tetrahymena provides strong support for the RNA splicing model which was developed.

TL;DR: High-specific-activity single-stranded (ss) [alpha-32P]DNA of a defined size complementary to sequences cloned into bacteriophage M13 is synthesized, which is up to 300 times more sensitive than conventional S1 mapping techniques.

TL;DR: Comparison of the sequences of human alpha-amylase cDNAs with those previously obtained for mouse alpha-AMylase genes showed the possibility of gene conversion between the two genes of humanalpha-amymylase.

TL;DR: The sequence of the 390 amino acid residues comprising the PMI monomer has been deduced, and the predicted Mr of 42 716 agrees with that for the protein of Mr 42 000 identified previously by the maxicell procedure.

TL;DR: Two families of plasmids suitable for the cloning of genes and for directing the synthesis of large amounts of fused proteins in Escherichia coli are constructed, which yields more than twice the amount produced by using a 1007 amino acid-long beta-galactosidase gene for fusion.

TL;DR: An improved bacteriophage λ cloning vector, λ2001, has been constructed that includes a 34-bp Polylinker oligonucleotide which introduces cleavage sites for Xba I, Sst I, Xho I, Eco RI, Hind III and Bam HI, and can accommodate 10-kb to 23-kb fragments.

TL;DR: Using a multicopy plasmid in which the tac promoter has been placed in front of the dam gene of Escherichia coli K-12, it is shown that levels of DNA adenine methylase activity are correlated with the spontaneous mutation frequency.