DNase I activity in pig MII oocytes: implications in transgenesis

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DNase I activity in pig MII oocytes: implications in transgenesis.

Several reliable methods to produce transgenic animals utilize the male genome. After penetration into oocyte, sperm DNA undergoes dramatic conformational changes that could represent a great opportunity for exogenous DNA to be integrated in the zygote genome. Among the enzymes responsible for sperm remodeling, a nuclease could be involved. The presence of a DNase I in oocytes has not been much...

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Animal models, particularly pigs, have come to play an important role in translational biomedical research. There have been many pig models with genetically modifications via somatic cell nuclear transfer (SCNT). However, because most transgenic pigs have been produced by random integration to date, the necessity for more exact gene-mutated models using recombinase based conditional gene expres...

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BACKGROUND The use of immature oocytes is limited to cases where these are the only available oocytes, and they are usually only microinjected with sperm after having undergone maturation in vitro. This study compares the outcome of injection of sperm into metaphase I oocytes immediately after their denudation (MI) performed 2 h after their retrieval, with the outcome of injection of sperm into...

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DNase I activity retained after heat inactivation in standard buffer.

For the detection of RNA transcripts by RT-PCR, prior removal of genomic DNA must be performed. To remove genomic DNA, RNA is often prepared by DNase I digestion following phenol extraction. Recently, one-tube or onebuffer systems of RT-PCR were developed to prevent loss of RNA and to reduce the risk of contamination (2,6). In these methods, DNase I is added before RT. Taq DNA polymerase prepar...

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ژورنال

عنوان ژورنال: Reproduction

سال: 2006

ISSN: 1470-1626,1741-7899

DOI: 10.1530/rep.1.00877