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Electroporation of abalone sperm enhances sperm-DNA association
Authors:F. Y. T. Sin,U. K. Mukherjee,J. C. McKenzie, I. L. Sin,&dagger  
Affiliation:Department of Zoology, University of Canterbury, Private Bag 4800, Christchurch 1, New Zealand;Department of Obstetrics and Gynaecology, Christchurch School of Medicine, Christchurch, New Zealand
Abstract:The ability of sperm from the black-footed abalone Haliotis iris to take up foreign DNA in solution has been demonstrated. The efficiency of DNA uptake is related to the conditions of electroporation, including field strength (625 V cm−1, 1000 V cm−1), pulse length (18.6 ms, 27.4ms) and number of pulses (1, 2), and DNA concentration (20, 100 μg ml−1). Sperm motility decreased with increased field strength and pulse number. At a field strength of 625 V cm−1, neither the pulse length nor pulse number enhanced DNA uptake. A 40% enhancement in DNA uptake was observed when the sperm were shocked at 1000 V cm−1 with two long pulses (27.4 ms each). Linear regression analysis revealed that pulse number ( p = 0.013) and field strength ( P =0.039) were the most important factors in sperm–DNA interaction. Higher DNA concentration enhanced sperm DNA uptake irrespective of field strength, pulse length and pulse number. The optimal electroporation conditions for DNA uptake were 1000 V cm', with two pulses of 27.4 ms each, and a DNA concentration of 100 μg ml−1.
Keywords:Haliotis iris    electroporation    sperm DNA uptake    gene transfer
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