The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow

Total genomic DNA from 24 normal cows and a normal male cattle were isolated from leucocyte preparation. Polymerase Chain Reaction (PCR) amplification of these genomic DNA using the PI-5EZ and P2-3EZ primers for ZFX/ZFY gene resulted in a fragment of approximately 440 bp in size from both male an...

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Main Author:	Shamsuddin, Shaharum
Format:	Thesis
Language:	English English
Published:	1996
Subjects:	Zinc-finger proteins - Research Y chromosome
Online Access:	http://psasir.upm.edu.my/id/eprint/8615/1/FSAS_1996_16_A.pdf
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spelling	my-upm-ir.86152012-05-08T04:33:25Z The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow 1996 Shamsuddin, Shaharum Total genomic DNA from 24 normal cows and a normal male cattle were isolated from leucocyte preparation. Polymerase Chain Reaction (PCR) amplification of these genomic DNA using the PI-5EZ and P2-3EZ primers for ZFX/ZFY gene resulted in a fragment of approximately 440 bp in size from both male and female samples. This PCR product represents part of the ZFX/ZFY gene defined by the primers used. The PCR products were proven to be amplified from bovine genomic DNA from the dot blot hybridization studies. After Pst I digestion, the female samples (ZFX) remained undigested while the male (ZFX/ZFY) showed three fragments of approximately 440 bp, 340 bp and 100 bp. Thus the ZFY gene has a unique Pst I site. The PCR product from a population of 45 female and 47 male cattle was further tested for the presence of the Pst I cleavage. There was no Pst I site in all the female (ZFX) samples, while 45 out of 47 samples from the male were cleaved. From X2 analysis of the data, the Pst I cleavage on ZFY was significantly sex dependent. The restriction analysis of ZFX/ZFY gene showed the existence of a unique Mnl I site on ZFX and more than one Alu I restriction site on both ZFX and ZFY. The PCR product from an infertile female cattle (supposedly, 60, XX and therefore ZFX/ZFX) was subjected to the Pst I digestion, and found to be cleaved indicating the presence of the ZFY gene and possibly the Y chromosome. The latter was confirmed by cytogenetic analysis. Post mortem and autopsy evaluation of the animal showed a partial development of male characteristic and a defective female reproduction system The use of the peR diagnosis for the presence of the Y chromosome was applied to a population of 30 suspected infertile female animals, and two were found to have the ZFY gene indicating the possible presence of the Y chromosome. Thus, peR amplification of the ZFX/ZFY gene followed by Pst I digestion and analysis on gel electrophoresis can be used as an approach to establish the presence of the Y chromosome in an infertile female cattle. This approach however, needs to be supported by karyo typing to establish unequivocally the screening procedure. Zinc-finger proteins - Research Y chromosome 1996 Thesis http://psasir.upm.edu.my/id/eprint/8615/ http://psasir.upm.edu.my/id/eprint/8615/1/FSAS_1996_16_A.pdf application/pdf en public masters Universiti Pertanian Malaysia Zinc-finger proteins - Research Y chromosome Faculty of Environmental Studies English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Zinc-finger proteins - Research Y chromosome
spellingShingle	Zinc-finger proteins - Research Y chromosome Shamsuddin, Shaharum The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
description	Total genomic DNA from 24 normal cows and a normal male cattle were isolated from leucocyte preparation. Polymerase Chain Reaction (PCR) amplification of these genomic DNA using the PI-5EZ and P2-3EZ primers for ZFX/ZFY gene resulted in a fragment of approximately 440 bp in size from both male and female samples. This PCR product represents part of the ZFX/ZFY gene defined by the primers used. The PCR products were proven to be amplified from bovine genomic DNA from the dot blot hybridization studies. After Pst I digestion, the female samples (ZFX) remained undigested while the male (ZFX/ZFY) showed three fragments of approximately 440 bp, 340 bp and 100 bp. Thus the ZFY gene has a unique Pst I site. The PCR product from a population of 45 female and 47 male cattle was further tested for the presence of the Pst I cleavage. There was no Pst I site in all the female (ZFX) samples, while 45 out of 47 samples from the male were cleaved. From X2 analysis of the data, the Pst I cleavage on ZFY was significantly sex dependent. The restriction analysis of ZFX/ZFY gene showed the existence of a unique Mnl I site on ZFX and more than one Alu I restriction site on both ZFX and ZFY. The PCR product from an infertile female cattle (supposedly, 60, XX and therefore ZFX/ZFX) was subjected to the Pst I digestion, and found to be cleaved indicating the presence of the ZFY gene and possibly the Y chromosome. The latter was confirmed by cytogenetic analysis. Post mortem and autopsy evaluation of the animal showed a partial development of male characteristic and a defective female reproduction system The use of the peR diagnosis for the presence of the Y chromosome was applied to a population of 30 suspected infertile female animals, and two were found to have the ZFY gene indicating the possible presence of the Y chromosome. Thus, peR amplification of the ZFX/ZFY gene followed by Pst I digestion and analysis on gel electrophoresis can be used as an approach to establish the presence of the Y chromosome in an infertile female cattle. This approach however, needs to be supported by karyo typing to establish unequivocally the screening procedure.
format	Thesis
qualification_level	Master's degree
author	Shamsuddin, Shaharum
author_facet	Shamsuddin, Shaharum
author_sort	Shamsuddin, Shaharum
title	The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
title_short	The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
title_full	The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
title_fullStr	The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
title_full_unstemmed	The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow
title_sort	use of zinc finger protein gene to detect the presence of y chromosome in the cow
granting_institution	Universiti Pertanian Malaysia
granting_department	Faculty of Environmental Studies
publishDate	1996
url	http://psasir.upm.edu.my/id/eprint/8615/1/FSAS_1996_16_A.pdf
_version_	1747810839655612416

The Use of Zinc Finger Protein Gene to Detect the Presence of Y Chromosome in the Cow

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