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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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 |
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English English |
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Zinc-finger proteins - Research Y chromosome |
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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 |