Showing papers in "Gene in 1991"

TL;DR: The results showed that high concentrations of G418 efficiently yielded L cell and CHO cell transfectants stably producing IL-2 at levels comparable with those previously attained using gene amplification.

TL;DR: Using the polymerase chain reaction and standard recombinant DNA techniques, a series of new multipurpose low-copy-number plasmids have been constructed, very useful for analyzing genes encoding proteins which are toxic in Escherichia coli in high copy number.

TL;DR: A new suicide vector (pKNG101) that facilitates the positive selection of double recombination events in Gram-bacteria has been developed and was used to mutate the blaA gene of Yersinia enterocolitica, by marker-exchange mutagenesis.

TL;DR: These studies demonstrate that the error rate per nucleotide induced in the 182 known detectable sites of the lacI gene was 1.6 x 10(-6) for Pfu DNA polymerase, a greater than tenfold improvement over the 2.0 x 10 (-5) error rate for Taq DNA polymerases.

TL;DR: Four new Escherichia coli cloning vectors are described, pUC6S, p UC21, pUK21 and pOK12, which contain a polylinker or multiple cloning site (MCS) with the recognition sequences for 28 restriction enzymes allowing blue/white screening for inserts.

TL;DR: A series of controlled expression vectors was constructed based on the wide-host-range plasmid pMMB66EH, finding that the tac promoter is five times stronger than the taclac promoter and that both were tightly regulated.

TL;DR: A family of cloning vectors derived from plasmid pACYC184 and, therefore, compatible with pBR322 and its derivatives (especially the pUC family of vectors), is described, which all contain a multiple cloning site (MCS) and the lacZ alpha reporter gene for easy cloning.

TL;DR: Two of the reporter enzymes most commonly used in studies of eukaryotic gene expression are chloramphenicol acetyl-transferase (CAT) and firefly luciferase (Luc) and more than tenfold increases in Luc activity can be observed when compounds that resemble its natural substrate, luciferin, are added.

TL;DR: Comparison between gene expression level and vector copy-number indicated that a plateau in delta-endotoxin production is reached with a copy- number of about fifteen per equivalent chromosome.

TL;DR: Two new broad-host-range plasmid vectors, p UCP18 and pUCP19, which are stably maintained in Escherichia coli and Pseudomonas aeruginosa have been constructed.

TL;DR: To increase the level of secreted mEGF, a synthetic secretion cassette encoding the alpha-factor prepro leader peptide from Saccharomyces cerevisiae fused to mouse epidermal growth factor was constructed and a method for rapidly screening large numbers of P. pastoris transformants for the presence of many copies of a foreign gene was developed.

TL;DR: A system is described for the single-copy, stable insertion of cloned DNA sequences into the chromosomes of Gram- bacteria using a narrow-host-range plasmid that contains the mini Tn7-Km transposon.

TL;DR: A phagemid that expresses a single-chain Ab fused to pIII, a coliphage protein product of gene III that initiates infection by binding to F pili is constructed, which can be tightly repressed so that Ab libraries can be amplified without risk of being dominated by deletion mutants.

TL;DR: The pVT-Bac, an Autographa californica nuclear polyhedrosis virus transfer vector that may enhance the secretion of other foreign proteins from insect cells as mentioned in this paper.

TL;DR: A gene, pilT, within the twitching motility region is predicted to encode a 344-amino acid protein which has strong homology to a variety of other bacterial proteins, which suggest that these are all related members of a specialised protein export pathway which is widespread in the eubacteria.

TL;DR: Many unique applications of this class of enzymes are described, including: precise trimming of DNA; retrieval of cloned fragments; gene assembly; use as a universal restriction enzyme; cleavage of single-stranded DNA; detection of point mutations; tandem amplification; printing-amplification reaction; and localization of methylated bases.

TL;DR: A new family of vectors including cloning vectors, an expression vector, and promoter probe vectors, has been constructed, based on the replication origins of the corynebacterial pBL1 and the Escherichia coli ColE1 plasmids, able to replicate in Corynebacterium glutamicum and E. coli.

TL;DR: Single-copy integration vectors suitable for cloning in S. aureus have been constructed based on the site-specific recombination system of staphylococcal phage, L54a, and the gene dosage of the DNA cloned in the vector matches that of the chromosome.

TL;DR: There is a compositional gradient of gene concentration in the human genome and the correlations for 'localized genes' and genes from the bank are in full agreement, indicating that the former set is representative of the latter.

TL;DR: This paper describes the protocol for the site-directed mutagenesis and an example of the introduction of several mutations in the hen egg-white lysozyme-encoding gene.

TL;DR: It is likely that a 'Klenow-like core', containing the DNA polymerase and 3'-5' exonuclease activities, has evolved from a common ancestor, giving rise to the present-day prokaryotic and eukaryotic DNA polymerases.

TL;DR: A plasmid was reisolated which transformed A. nidulans at a frequency of 20,000 transformants per 10(6) protoplasts at near saturation levels of transforming DNA, representing a 250-fold enhancement of transformation efficiency over that found for typical integrative vectors such as pILJ16, theplasmid used in gene bank constructions.

TL;DR: The mobilizable shuttle cloning vectors, pAT18 and pAT19, are composed of ten unique cloning sites that allow screening of derivatives containing DNA inserts by alpha-complementation in Escherichia coli carrying the lacZ delta M15 deletion and can be efficiently mobilized by self-transferable IncP plasmids co-resident in the E. coli donors.

TL;DR: Low-usage codons are clearly avoided in genes encoding abundant proteins for ECO, YSC DRO, and several species of primates, with an emphasis on characterization of low- usage codons.

TL;DR: The isolation of a phage lambda genomic clone of the murine Spp-1 gene containing the promoter and first six exons is reported, and another exon located 5' to the 'exon 1' reported by Miyazaki et al is found.

TL;DR: Rec recombinant adenoviruses, with functional or defective E1a genes, which harbor either the hepatitis B (HB) virus gene encoding the HB surface antigen, as well as the pre-S2 epitopes, or the bacterial gene encoding chloramphenicol acetyltransferase (CAT) under control of the Ad major late promoter (MLP).

TL;DR: The nature and properties of a variety of plasmids are described that facilitate the construction of baculovirus vectors for expression of one or more heterologous genes, designed for use with Autographa californica nuclear polyhedrosis virus, AcMNPV, as a vector for protein production in insect cells and/or insect larvae.

TL;DR: The entire nucleotide (nt) sequences of three genes (xlnA, xlnB and xlnC) of Streptomyces lividans encoding three distinct xylanases (Xln) have been determined and showed some homology with either the N- termini or the C-termini of eight other Xln and of two exo-glucanases.

TL;DR: It is found that the gene conferring resistance to puromycin from Streptomyces alboniger does not influence adjacent promoters, and the neo gene is frequently used in eukaryotic vectors as a dominant selectable gene.

TL;DR: The XYL1 gene of the yeast Pichia stipitis has been isolated from a genomic library using a specific cDNA probe, and its nucleotide sequence has been determined.