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University of Nebraska - Lincoln

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Articles 121 - 125 of 125

Full-Text Articles in Biochemical and Biomolecular Engineering

Effects Of Time Of Deoxyribonucleic Acid Microinjection On Gene Detection And In Vitro Development Of Bovine Embryos, A.E T. Sparks, R S. Canseco, C G. Russell, J L. Johnson, H D. Moll, William H. Velander, Francis C. Gwazdauskas Mar 1994

Effects Of Time Of Deoxyribonucleic Acid Microinjection On Gene Detection And In Vitro Development Of Bovine Embryos, A.E T. Sparks, R S. Canseco, C G. Russell, J L. Johnson, H D. Moll, William H. Velander, Francis C. Gwazdauskas

Papers in Biotechnology

In vivo fertilized embryos were surgically collected from superovulated dairy cows to evaluate microinjection on embryo development and utilized the polymerase chain reaction technique for selection of transgenic embryos. Seventy-two percent of the embryos with visible pronuclei or nuclei were microinjected with DNA, and the remaining 28% served as uninjected controls. All embryos were cocultured with bovine oviductal epithelial cells. Mean final development scores of embryos within the same initial cell stage at collection were unaffected by microinjection. After 144 h of culture, 45% of the microinjected embryos developed to the morula or blastocyst stage. The transgene was detected in …


High-Level Expression Of A Heterologous Protein In The Milk Of Transgenic Swine Using The Cdna Encoding Human Protein C, William H. Velander, John L. Johnson, Raymond L. Page, Chritopher G. Russell, Anuradha Subramanian, Tracy D. Wilkins, Francis C. Gwazdauskas, Christoph Pittus, William N. Drohan Dec 1992

High-Level Expression Of A Heterologous Protein In The Milk Of Transgenic Swine Using The Cdna Encoding Human Protein C, William H. Velander, John L. Johnson, Raymond L. Page, Chritopher G. Russell, Anuradha Subramanian, Tracy D. Wilkins, Francis C. Gwazdauskas, Christoph Pittus, William N. Drohan

Papers in Biotechnology

Transgenic pigs were generated that produced human protein C in their milk at up to 1 g/liter. The gene construct was a fusion gene consisting of the cDNA for human protein C inserted into the first exon of the mouse whey acidic protein gene. These results demonstrate that the mouse whey acidic protein gene contains regulatory elements that can direct cDNA expression at high levels in the pig mammary gland. Recombinant human protein C that was produced at about 380 pg/ml per hr in transgenic pig milk possessed anticoagulant activity that was equivalent to that of protein C derived from …


In Vitro Development Of Zygotes From Prepubertal Gilts After Microinjection Of Dna, B. L. Williams, J. L. Johnson, A. E.T. Sparks, R. S. Canseco, J. W. Knight, William H. Velander, R. L. Page, W. N. Drohan, J. M. Young, R. E. Pearson, T. D. Wilkins, F. C. Gwazdauskas Jul 1992

In Vitro Development Of Zygotes From Prepubertal Gilts After Microinjection Of Dna, B. L. Williams, J. L. Johnson, A. E.T. Sparks, R. S. Canseco, J. W. Knight, William H. Velander, R. L. Page, W. N. Drohan, J. M. Young, R. E. Pearson, T. D. Wilkins, F. C. Gwazdauskas

Papers in Biotechnology

The effect of pronuclear microinjection of DNA and culture in excised mouse oviducts on the development of porcine zygotes was assessed in this study. Precocious ovulation was induced in prepubertal gilts with pregnant mare's serum gonadotrophin and hCG. Zygotes received either pronuclear microinjection of buffer alone, buffer containing a DNA construct, or no microinjection. Zygotes were cultured in vitro in either modified Krebs-Ringer bicarbonate medium (KRBI for 144 h or in mouse oviduct (MO) explant culture with KRB for 48, 72, 96, or 120 h. Pronuclear microinjection of DNA resulted in a lower (P < .05) cleavage index (CII than did buffer or no microinjection (CI 2.16 k .10 vs 2.80 f .13 and 2.93 f .lo). The CI loss was greatest for DNA-injected zygotes at the two-cell stage of development. Coculture of zygotes in MO resulted in a higher CI (P < .01) than did culture in KRB. Culture in MO for 72 h was the most beneficial system compared with MO for 48, 96, or 120 h (P < .05; CI 3.25 k .12 vs 2.66 k .18, 2.79 f .14, and 2.40 .14, respectively). Microinjection of DNA, not merely the mechanical procedure, was detrimental to early zygote development and may be the cause of low pregnancy rates.


Enzyme- Facilitated Transport And Separation Of Organic Acids Through Liquid Membranes, David C. Rethwisch, Anuradha Subramanian, Gao Yi, Jonathan S. Dordick Jan 1990

Enzyme- Facilitated Transport And Separation Of Organic Acids Through Liquid Membranes, David C. Rethwisch, Anuradha Subramanian, Gao Yi, Jonathan S. Dordick

Papers in Biotechnology

The selective separation of organic acids is important for the production of numerous biologially functional rnuleculcs, including amino acids, peptidcs, fats, and pharmaceuticals.' Enzyme, specificalty lipases and proteases, have been used to selectively separate a variety of organic acids; however, multiple steps are usually required,Vn additiun, carbonic anhydrase has been usd in a variety of Eiquid membranes for transport of CO2.) In the present study, we have coupled the selectiviey of enzymes with liquid membranes to provide a single-step method to scicctiveiy separate and purify organic acids.


Conformational Changes In An Epitope Localized To The Nh2-Terminal Region Of Protein C, Carolyn L. Orthener, Rapti D. Madurawe, William H. Velander, William N. Drohan, Frances D. Bettey, Dudley K. Strickhand Nov 1989

Conformational Changes In An Epitope Localized To The Nh2-Terminal Region Of Protein C, Carolyn L. Orthener, Rapti D. Madurawe, William H. Velander, William N. Drohan, Frances D. Bettey, Dudley K. Strickhand

Papers in Biotechnology

Murine monoclonal antibodies, developed following immunization with human protein C, were characterized for their ability tboin d antigen in thpe resence of either CaClz or excess EDTA. Three stablec lones were obtained which produced antiboditehsa t bound to protein C only itnh e presence of EDTA. Allt hree antibodies bound to the light choafi np rotein C on immunoblots and also bound to the homologous proteins factor X and prothrombin in solid-phase radioimmunoassays. One antibody, 7D7B10 was purified and studied further. The binding of 7D7B10 to human protein C was characterized bya KOo f 1.4 nM. In competition studies, it …