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A Method For The Large Scale Purification Of Hcg (Human Chorionic Gonadotropin) From The Urine Of Pregnant Women, Chun-Shyan Kuo

Chemistry & Biochemistry Theses & Dissertations

A large scale preparation of human chorionic gonadotropin (HCG) from the urine of pregnant women was accomplished by using kaolin adsorption, NH₄OH extraction and DEAE-cellulose ion exchange. A recovery about 81% of HCG and a specific activity in the 2,000-3,000 IU/mg range was achieved using this purification method.

Two HCG activity fractions, HCG I & HCG II, were isolated from DEAE-cellulose chromatography by a stepwise sodium chloride gradient elution. HCG I was eluted at 0.01-0.02 M NaCl while HCG II was eluted at 0.12-0.02 M NaCl.

The differences between HCG I and HCG II have been studied. The …

Modulation Of Queuine Uptake In Cultured Human Fibroblasts By Phorbol Esters And Interferons, Debra L. Crane

Chemistry & Biochemistry Theses & Dissertations

Queuosine, a nucleoside found exclusively in the first position of the anticodon of transfer RNA (tRNA), is generated post-transcriptionally in an exchange of guanine for queuine by the modifying enzyme tRNA-guanine ribosyltransferase. Phorbol ester tumor promoters were shown to induce queuine hypomodified tRNA, and that phorbol ester action was due, in part, to inhibition of queuine transport across the cell membrane. An unidentified protein designated conditioned medium factor (CMF) that potentiated a phorbol-induced inhibition of queuine uptake was also documented. We suggest here that phorbol ester-induced inhibition of queuine uptake is not a significant factor in inducing queuine-deficient tRNA although …

Molecular Study Of The B19 (Human) Pathogenic Parvovirus, Jamshed Ayub

Theses and Dissertations in Biomedical Sciences

The B19 (human) parvovirus is a small single stranded DNA virus of 5.4 kilobases. B19 is specific for erythroid progenitor cells and has been propagated in vitro only with human erythroid bone marrow. Replication of viral DNA and the viral protein products of B19 appear similar to those of other animal parvoviruses. However, B19 differs from other parvoviruses in some important aspects, which include the initiation of all transcripts at a strong left side promoter (p6) and the absence of a functional internal promoter. B19 has an unusual transcription map which is described in this study.

The transcription map of …

Biochemical And Histological Evaluation Of Normal Human Epidermis Vs Cultured Human Epidermis, Ngo Thi Bich Lien

Biological Sciences Theses & Dissertations

Cultured human keratinocytes were grown in vitro for transplantation purposes in case of severe burns. Single keratinocytes were harvested from human epidermis and grown in culture flasks such that multilayered sheets of keratinocyte cells were harvestable in approximately three weeks. In order to evaluate cultured epidermis for equivalency with normal human epidermis, biochemical and histological studies were performed. Thin sections of cultured epidermis were shown to be composed of cells that had undergone developmental differentiation, but not terminal differentiation. Biochemical analyses of keratin and prekeratin proteins isolated from cultured epidermis indicated that cultured keratinocytes have electrophoretic patterns similar to keratins/prekeratins …

Yeast Mannans: Use As A Transport Vehicle For Peptides, Jacqueline Cesario Lee

Masters Theses

Yeast mannans are phosphorylated glycopeptides found in various species of yeast. Their similarity to the mannose-phosphate signal for intracellular lysosomal uptake led mannans to be considered as a possible vehicle for increasing cellular affinity for peptides. Insulin was used as a model peptide and was conjugated to mannans by two coupling methods. The conjugated mannan-insulins were compared to insulin on the basis of cellular binding and ability to induce typical insulin biological effects.

Solid Phase Extraction Of Mammalian Cell Mitochondrial Dna And Its Electrophoretic Separation In Agarose Gels, Theresa Brick-Miller

Biological Sciences Theses & Dissertations

An improved and efficient electrophoretic procedure for the isolation of mitochondrial DNA (mtDNA) is described. The solid phase extraction procedure for the isolation of mtDNA involves embedding as few as 200,000 Ehrlich ascites tumor cells into a block of agarose, digestion of cellular membranes by detergent action and subsequent electrophoretic separation of nucleic acids in agarose gels. Genomic DNA remains in the original block of agarose while the mtDNA migrates in the separation gel to a position equivalent to that obtained for mtDNA isolated by traditional procedures.

The efficacy of a large number of detergents, agarose concentrations, embedding conditions, embedding …

Isolation And Characterization Of Temperature-Sensitive Protein Synthesis Mutants Of Escherichia Coli By Directed Mutagenesis Of The Defective Bacteriophage Lambda Fus2, Kenton L. Lohman

Electronic Theses and Dissertations

Mutagenesis of the defective transducing bacteriophage lambda fus2 was used to isolate a collection of temperature-sensitive mutants of E. coli in the major ribosomal protein gene cluster. Four mutants were examined in detail. Two of the mutants were resistant to the ribosomal antibiotics neamine and spectinomycin. Another mutant was defective in 50S ribosomal subunit assembly at 42(DEGREES)C. The 30S subunit proteins S17 and S19 were changed in two different mutants. Each protein migrated as a more basic species in two-dimensional gels of ribosomal proteins. Ribosomes from each of the four mutants examined showed a temperature-dependent reduction in translational activity in …

Lac Operon Expression In Steady State Cells Of Escherichia Coli, Sungae Cho

Dissertations, Theses, and Masters Projects

No abstract provided.

Characterization Of Kpni Interspersed, Repetitive Dna Sequence Families And Their Association With The Nuclear Matrix, Joseph A. Chimera

Electronic Theses and Dissertations

The KpnI, 1.2 and 1.5 kb families of interspersed repetitive DNAs from the African green monkey genome were isolated and characterized. Each family contains three populations of segments based on their sequence lengths and susceptibility to cleavage by the restriction enzymes KpnI and RsaI. The first population contains the smallest segments which are susceptible to both KpnI and RsaI cleavage and have fragment lengths of 1.2 kb (1.2 kb family) and 1.5 kb (1.5 kb family) respectively. The members in this population are referred to as KpnI-sensitive segments. The second population contains longer segments (> 2 kb) which represent fusions …

Comparison Of Surface Polysaccharides Of R. Leguminosarum Anu57 (Exo-1), R. Leg. Anu57(Pbr1an) (Nod+, Nif-), And R. Leg. Anu57(Pjb5j1) (Nod+, Nif-), Frank G. Hustmyer

Masters Theses

Extracellular polysaccharides (EPS) and lipopolysaccharides (LPS) are suspected to be involved in the legume/Rhizobium symbiotic process. Polysaccharide compositions from the EPS, LPS, capsular polysaccharide (CPS), and small polysaccharide (SmPS) were compared between Rhizobium leguminosarum ANU57 (parent, Exo^-1) and two mutants R. leg. ANU57(pBR1AN) (nod⁺, nif^-) and R. leg. ANU57(pJB5J1) (nod⁺, nif^-) both containing a Sym (symbiotic) plasmid insertion. R. leg. ANU57(pBR1AN) and R. leg. ANU57(pJB5J1) are nod⁺ on clover and peas, respectively. The nod⁺ mutants produce larger amounts of EPS and decreased amounts of LPS compared to …

Determination Of Erythrocyte Zinc Concentrations Using Zeeman Effect Atomic Absorption Spectroscopy, Tanya Maranda Lewis

Chemistry & Biochemistry Theses & Dissertations

The purpose of this research project was to develop assay procedures for determination of zinc in erythrocytes. Three methodologies were developed. The methods were evaluated on the basis of within-run and between-run precision. The most reproducible method, the 213.856 nm flame methodology, was used to determine red blood cell zinc concentrations in more than 200 samples obtained from the Neonatal Intensive care Unit at the Children's Hospital of the King's Daughters.

This study demonstrated that erythrocyte zinc levels in premature infants were similar to those of healthy adults. A linear relation between plasma and red blood cell zinc concentrations was …

Isomers Of 5.8s Ribosomal Rna, David Smith

Chemistry & Biochemistry Theses & Dissertations

When whole cell RNA was separated by high resolution PAG (i.e., polyacrylamide gel) electrophoresis, 5.8S rRNA migrated as three higher mobility intense bands, and three faint bands of lower mobility. The three intense bands represent conformational isomers which migrated as a single band under denaturing conditions on PAG. The three faint bands represent conformational isomers of a minor form of 5.8S rRNA which is elongated at the 5' terminus of the molecule. This minor form migrates as a single band of lower mobility than the major form when electrophoresed on a denaturing PAG. The minor form was determined to have …

Functions Of Deadenylation Factors In Mrna Decay And Mrna Processing Body Formation, Dinghai Zheng

Dissertations & Theses (Open Access)

Most newly synthesized messenger RNAs possess a 5’ cap and a 3’ poly(A) tail. The process of poly(A) tail shortening, also termed deadenylation, is important for post-transcriptional gene regulation, because deadenylation not only leads to mRNA translational inhibition but also is the first step of major mRNA degradation. Translationally inhibited mRNAs can be stored and/or degraded in dynamic cytoplasmic foci termed mRNA processing bodies, or P bodies, which are conserved in eukaryotes.

To shed new light on the mechanisms of P body formation and P body functions, I focused on the link between deadenylation factors and P bodies. I found …

High Resolution Two-Dimensional Electrophoretic Study Of Human Seminal Plasma Proteins, Edward E. Gaunt

Chemistry & Biochemistry Theses & Dissertations

A comprehensive biochemical screening procedure for the evaluation of seminal plasma constituents is needed which, when used in conjunction with semen analysis, can provide the physician with both qualitative and quantitative information regarding the fertility status of an individual.

High resolution two-dimensional electrophoresis is a technique whereby proteins in a complex mixture such as a biological fluid are separated in one dimension by molecular charge and in the second dimension by molecular mass. It has been the purpose of this investigation to adapt and evaluate this technique for the analysis of human seminal plasma.

Our efforts have shown that this …

The Association Of 5.8 S With 28s Ribosomal Rna, Nandita Banerjee

Chemistry & Biochemistry Theses & Dissertations

5.8S rRNA is a low molecular weight ribosomal RNA which is noncovalently bonded to the larger ribosomal subunit 28S rRNA; through its 3' end and through its 5' end. This interaction is an integral part of the ribosome, and plays an important role in the ribosome structure and function.

There is a high degree of homology between the 5.8S rRNA primary structures of rat, turtle and chicken. The base sequence of rat 5.8S rRNA differs only in one position from that of turtle and in three positions fr.om that of cl1.icken. Tl1ere is a single purine substitution at the 5' …

Multiple Methods Of Analysis Of 2'-O-Methylation In 5.8s Rrna, Sharon H. Ryan

Chemistry & Biochemistry Theses & Dissertations

5.8S rRNA is a low molecular weight ribosomal RNA which is found hydrogen bonded to 28S rRNA in the eucaryotic cell. There are two nucleotides which have 2'-0-methylations; a guanine residue at position 77 is fully methylated and a uridine residue at position 14 which is partially methylated. This partial 2'-0-methylation of the uridine residue has been found to vary with the tissue source, with the highest level in normal tissue and the lowest level in neoplastic tissue. In order to study the significance of this site-specific methylation, a reliable and convenient method of analysis was needed.

Early studies on …

Purification And Characterization Of Dihydroorotase From Clostridium Oroticum, A Zinc-Containing Metalloenzyme, William Edward Balch

Dissertations and Theses

Dihydroorotase (4,5-L-dihydro-orotate amidohydrolase, EC 3.5.2.3) which catalyzes the reversible cyclization of N-carbamyl-1-aspartate toL-dihydro-orotate has been purified from orotate-grown Clostridium oroticum by a combination of streptomycin sulfate fractionation, DEAE-Sephadex chromatography, and hydroxylapatite chromatograpy. The enzyme has been shown to be omogeneous when subjected to polyacrylamide gel electrophoresis. Thin-layer gel chromatography with Sephadex G-200 indicated the enzyme to have a molecular weight of 110,000 ± 10,000. Sodium dodecyl sulfate gel electrophoresis using two different buffer systems indicate the enzyme to be composed of two identical subunits with a molecular weight of 56,000 ± 5300. Dihydroorotase has been shown to be a zinc-containing …

Membrane Bound Nadh Oxidase Complex Of Micrococcus Lysodeikticus, Karen Ju-Kuang Hsia

Masters Theses

No abstract provided.

The Pattern Of Rna Synthesis In Dormant And Germinating Statoblasts Of Pectinatella Magnifica, Virginia Kent Proud

Dissertations, Theses, and Masters Projects

No abstract provided.

A Comparison Of Ribosomal Ribonucleic Acid Using Disc Electrophoresis, Carolyn Gene Mohler

Dissertations, Theses, and Masters Projects

No abstract provided.