The binding of exogenous DNA fragments to bovine Spermatozoa

J. M. Bird; F. Gannon; R. Powell; R. Horan; J. A. Houghton

doi:10.1080/10495399209525772

The binding of exogenous DNA fragments to bovine Spermatozoa

J. M. Bird, F. Gannon, R. Powell, R. Horan, J. A. Houghton

Infectious Disease

University of Galway

Research output: Contribution to a Journal (Peer & Non Peer) › Article › peer-review

1 Citation (Scopus)

Abstract

The ability of mature, freeze-thawed bovine sperm to bind exogenous end-labelled or oligo-labelled X Hind III DNA restriction fragments was examined. Following 30 min. incubation of bovine sperm with P³²end-labelled A Hind III DNA and five washes with medium, approximately 5.8 ng DNA were bound to 10⁷sperm. Agarose gel autoradiography revealed that all of the λ Hind III DNA bands were present following sperm washes except for the smaller 0.5 Kb and 0.125 Kb bands. Incubation of sperm with ³H oligo-labelled λ Hind III DNA gave a much higher level of binding (138 ng/10⁷sperm) than that found with end-labelled DNA. This binding was entirely eliminated by DNase I. The separation of live and dead sperm fractions on Percoll gradients revealed that more oligo-labelled A Hind III DNA was found to be associated with the dead sperm fraction (31.2 ng/10⁷sperm) rather than the live sperm fraction (2.7 ng/10⁷sperm). Analysis of supravital stained, light microscopic autoradiographs confirmed that oligo-labelled λ Hind III DNA bound to dead sperm in the post-acrosomal region of the sperm head although other minor distribution patterns were observed.

Original language	English
Pages (from-to)	181-200
Number of pages	20
Journal	Animal Biotechnology
Volume	3
Issue number	2
DOIs	https://doi.org/10.1080/10495399209525772
Publication status	Published - 1 Jan 1992

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title = "The binding of exogenous DNA fragments to bovine Spermatozoa",

abstract = "The ability of mature, freeze-thawed bovine sperm to bind exogenous end-labelled or oligo-labelled X Hind III DNA restriction fragments was examined. Following 30 min. incubation of bovine sperm with P32end-labelled A Hind III DNA and five washes with medium, approximately 5.8 ng DNA were bound to 107sperm. Agarose gel autoradiography revealed that all of the λ Hind III DNA bands were present following sperm washes except for the smaller 0.5 Kb and 0.125 Kb bands. Incubation of sperm with 3H oligo-labelled λ Hind III DNA gave a much higher level of binding (138 ng/107sperm) than that found with end-labelled DNA. This binding was entirely eliminated by DNase I. The separation of live and dead sperm fractions on Percoll gradients revealed that more oligo-labelled A Hind III DNA was found to be associated with the dead sperm fraction (31.2 ng/107sperm) rather than the live sperm fraction (2.7 ng/107sperm). Analysis of supravital stained, light microscopic autoradiographs confirmed that oligo-labelled λ Hind III DNA bound to dead sperm in the post-acrosomal region of the sperm head although other minor distribution patterns were observed.",

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T1 - The binding of exogenous DNA fragments to bovine Spermatozoa

AU - Bird, J. M.

AU - Gannon, F.

AU - Powell, R.

AU - Horan, R.

AU - Houghton, J. A.

PY - 1992/1/1

Y1 - 1992/1/1

N2 - The ability of mature, freeze-thawed bovine sperm to bind exogenous end-labelled or oligo-labelled X Hind III DNA restriction fragments was examined. Following 30 min. incubation of bovine sperm with P32end-labelled A Hind III DNA and five washes with medium, approximately 5.8 ng DNA were bound to 107sperm. Agarose gel autoradiography revealed that all of the λ Hind III DNA bands were present following sperm washes except for the smaller 0.5 Kb and 0.125 Kb bands. Incubation of sperm with 3H oligo-labelled λ Hind III DNA gave a much higher level of binding (138 ng/107sperm) than that found with end-labelled DNA. This binding was entirely eliminated by DNase I. The separation of live and dead sperm fractions on Percoll gradients revealed that more oligo-labelled A Hind III DNA was found to be associated with the dead sperm fraction (31.2 ng/107sperm) rather than the live sperm fraction (2.7 ng/107sperm). Analysis of supravital stained, light microscopic autoradiographs confirmed that oligo-labelled λ Hind III DNA bound to dead sperm in the post-acrosomal region of the sperm head although other minor distribution patterns were observed.

AB - The ability of mature, freeze-thawed bovine sperm to bind exogenous end-labelled or oligo-labelled X Hind III DNA restriction fragments was examined. Following 30 min. incubation of bovine sperm with P32end-labelled A Hind III DNA and five washes with medium, approximately 5.8 ng DNA were bound to 107sperm. Agarose gel autoradiography revealed that all of the λ Hind III DNA bands were present following sperm washes except for the smaller 0.5 Kb and 0.125 Kb bands. Incubation of sperm with 3H oligo-labelled λ Hind III DNA gave a much higher level of binding (138 ng/107sperm) than that found with end-labelled DNA. This binding was entirely eliminated by DNase I. The separation of live and dead sperm fractions on Percoll gradients revealed that more oligo-labelled A Hind III DNA was found to be associated with the dead sperm fraction (31.2 ng/107sperm) rather than the live sperm fraction (2.7 ng/107sperm). Analysis of supravital stained, light microscopic autoradiographs confirmed that oligo-labelled λ Hind III DNA bound to dead sperm in the post-acrosomal region of the sperm head although other minor distribution patterns were observed.

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JO - Animal Biotechnology

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The binding of exogenous DNA fragments to bovine Spermatozoa

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