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T Boone

Publications and source records attributed to T Boone.

80 records · Page 5Linked to original sources
PubMed1978

Hybrid plasmids containing the araBAD genes of Escherichia coli B/r.

The DNA fragments generated by restriction endonuclease BamI which contain the araCBAD genes from E.coli B/r have been cloned. The DNA fragments containing ara genes were idenified by a compairson of the BamI fragments of lambdah80dara phages containing different ara deletion mutations. The ara genes were cloned into the plasmid pBR317, a derivative of ColE1. The cloned DNA fragments were analyzed by digestion with pairs of restriction endonucleases to determine the molecular weight of the chimeras and to identify the cloned ara DNA fragments. The cloned ara fragments were also identified by genetic complementation and recombination tests.

Arabinose↗
PubMed1978

DNA sequence of the araBAD promoter in Escherichia coli B/r.

The L-arabinose operon in Escherichia coli is a model system for the study of the control of gene expression. Maximal expression of the araBAD operon requires two positive control components: the araC protein-L-arabinose complex and the cyclic AMP receptor protein-cyclic AMP complex. Both araC protein and cyclic AMP receptor protein are required for the initiation of transcription of araBAD mRNA. We have used the plasmid pBR322 as a vector for cloning DNA fragments that contain the araBAD promoter. The cloned ara fragments were identified by both physical and genetic tests. A restriction map was constructed and the DNA sequence of the promoter was determined. The promoter contains a site that is similar to the RNA polymerase recognition sites in the galactose and lactose operons. It also contains a region similar to the known cyclic AMP receptor protein binding sites in the galactose and lactose operons.

Arabinose↗
PubMedDate not supplied

Identification and characterization of a nucleotide binding site on recombinant murine granulocyte/macrophage-colony stimulating factor.

Granulocyte/macrophage-colony stimulating factor (GM-CSF) is a regulatory cytokine important in the proliferative and functional activation of hematopoietic cells. It belongs to a family of 20 kDa or less acidic glycoprotein molecules found in a broad range of cellular sources. On the basis of the previously reported nucleotide-binding properties of interleukin-2 (IL-2), atrial natriuretic factor (ANF), and glucagon, the interaction of GM-CSF with nucleotides was investigated. Using radiolabeled 8-azidoadenosine-containing photoprobes of ATP ([gamma-32P]-8N3ATP) and Ap4A, the putative biological alarmone ([beta'-32P]-8N3Ap4A), we have identified a nucleotide binding site on recombinant murine GM-CSF (rmGM-CSF). Specificity of binding was demonstrated by saturation and competition experiments. Saturation of photoinsertion by [gamma-32P]-8N3ATP and [beta'-32P]-8N3Ap4A occurs with apparent Kd's of 10 and 0.7 microM, respectively. Using an immobilized Fe3+ affinity chromatography technique, developed specifically for the isolation of photolabeled peptides, a single radiolabeled peptide was isolated. It was identified as amino acids 5-14 near the N-terminus of GM-CSF. This peptide region has been shown in previous studies to be critical for biological activity. Also consistent with this observation is our finding that the photolabeled GM-CSF has lost most, if not all, of its biological activity, as determined by a cellular proliferation assay.

Adenosine Triphosphate↗
PubMedDate not supplied

Preparative electrofocusing for industrial applications.

This paper presents a case study of using a multicompartment isoelectric focusing apparatus to determine the isoelectric points and focus preparative quantities of brain derived neurotrophic factor (BDNF) and neurotrophic factor 3 (NT3). A separation of PEGylated from unPEGylated forms of granulocyte colony stimulating factor (G-CSF) is described as well. Both BDNF and NT3 have substantially higher pI values in this system than is predicted from sequenced based modeling. Although PEGylated forms of G-CSF can be separated from the unPEGylated forms, separation of protein with differing degrees of PEGylation was not achieved. The paper additionally demonstrates that this technique can be used simultaneously as an analytical and preparative tool, eliminating the need for analytical IEF gels.

Brain-Derived Neurotrophic Factor↗
PubMedDate not supplied

Evaluation of recombinant canine granulocyte colony-stimulating factor as an inducer of granulopoiesis. A pilot study.

Five healthy young adult dogs were given recombinant canine granulocyte colony-stimulating factor (rcG-CSF) at a dosage of 5 micrograms/kg/day subcutaneously for 4 weeks to evaluate the effect on complete blood cell counts. The mean neutrophil counts +/- standard deviation (SD) increased significantly (P less than 0.01) from 6,537/microliters +/- 1,726 (range, 4,950-9,512/microliters) to 26,330/microliters +/- 7,066 (range, 15,368-35,785/microliters) within 24 hours after the first injection of rcG-CSF. Mean monocyte counts +/- SD were significantly increased (P less than 0.05) from baseline values of 751/microliters +/- 168 (range, 444-891/microliters) to 2,514/microliters +/- 878 (range, 1,740-3,752/microliters) on day 5 of rcG-CSF administration. Mean neutrophil and monocyte counts (+/- SD) continued to increase reaching a maximum of 72,125/microliters +/- 15,073 (range, 50,915-96,278/microliters) and 3,972/microliters +/- 2,621 (range, 685-8,030/microliters), respectively by day 19. These increased neutrophil and monocyte counts were maintained until the administration of rcG-CSF was stopped. Blood counts returned to normal within 5 days after discontinuing the rcG-CSF. One week after discontinuing treatment, rcG-CSF was started again at 5 micrograms/kg/day subcutaneously. Within 48 hours following administration of rcG-CSF, mean neutrophil counts +/- SD increased from 5,860/microliters +/- 1,819 (range, 3,720-8,650/microliters) to 57,444/microliters +/- 8,173 (range, 43,983-68,278/microliters). Myeloid:erythroid ratios increased from a mean of 1.63:1 on day 1 prior to administration of rcG-CSF to 3.3:1 on day 10 in three dogs for which bone marrow samples were evaluated. Recombinant canine G-CSF did not cause clinically significant toxicosis in any of the dogs.

Animals↗