Administrative Information
)1321/1/7 ‫ تاريخ‬31-30-21(
‫المعلومات االدارية‬
Project Title - ‫عنوان المشروع‬
Optimization of genetic markers characterization in goat’s sperms: a prerequisite to future
selection strategy
‫ ببعص‬:‫كشجينيب يبا كشئببعالم كش نبما ش‬
‫ضبط رب ا كشف ب‬
‫صبئ ابعالم كشال ببع‬
‫ي‬:‫أمشمي تئبق كئت كتيجي كال تيع كش ئتقب‬
Principal Investigator - ‫الباحث الرئيسي‬
‫قم كشهعت‬
Telephone
001111-10
‫كشانمكئ كالشفت منا‬
e-mail
b.uduabl @al.b
‫كشانمكئ‬
Address
FAFS/AUB
‫كشمظيفي‬
Post
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‫كش ؤئئ‬
Institution
‫كالئم‬
Name
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Co-Workers - ‫الباحثون المشاركون‬
‫كشانمكئ كالشفت منا‬
e-mail
‫كش ؤئئ‬
Institution
‫كالئم‬
Name
ni.uduabl @al.b
A AhS Ur
hSu@K Hunu@ S
tSunu@ Sduabl @al.b
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Duration -‫المدة التعاقدية للمشروع‬
Scientific Information
‫العلمية‬
‫المعلومات‬
ّ
Objectives - ‫الهدف‬
The project purpose is to implement for the first time a project for optimization of protocols
needed in characterization of genes responsible for production, immunity, and susceptibility to
scrapie disease in Baladi goats of Lebanon, and to attempt to separate the X from the Y –
sperms of these males. This new project, will result in an indispensible data needed in the future
for sound selection programs to improve the productivity and immunity of the Baladi goats, and
to ensure in the selection process the establishment of goats flocks that are free of scrapie genes
that are responsible for inducing v-CJD in humans. In addition, the success in separation of X
and Y sperms, can predict in the future artificial insemination programs, the sex of the expected
newly born goats.
Achievements –‫أالنجازات المحققة‬
Selection for immunity improvement and prion scrapie control in goats, based on a respective
sperm-genetic markers polymorphism and nucleotide sequences, is concluded in Saanen and
Local breeds. Standardization of the amplification of the second exon of the caprine MHC
classII DRB gene, followed by PstI and TaqI restriction endonucleases reactions, confirmed the
polymorphism in this marker that could correlate with different levels of immunity during
selection of goats.
In addition, the amplification of the prion gene of goat sperms is standardized, revealing
nucleotide sequences that could be used as markers during selection for elimination of
susceptibility to scrapie disease.
Selection for production improvement in goats, based on specific sperm-genetic markers
polymorphism, is concluded. The amplified genetic markers that were subjected to restriction
endonuclease reactions included genes of the Growth Hormone and αS1-casein. Amplification
standardization was based on reading the concentration of the amplicons resulting from many
trials, while polymorphism was detected by images of the documented banded fragments
resulting from the endonucleases activities.
The significance of the polymorphism detected in genes of the Growth Hormone and that of
αS1-casein in correlation to production during the selection process is discussed.
Perspectives - ‫آفاق البحث‬
Standardization of goat sperm gene amplification is different for different genes, regardless of the use of
the same constant concentration of the goat sperm-DNA. The restriction endonuclease TaqI is successful
in revealing polymorphism in MHC class II gene of goats but not by the PstI digestion.
Sequencing of prion gene in local Lebanese goat sperms has to be done on freshly extracted DNA and
not the frozen-thawed substrate to reduce the reporting of unknown nucleotides. The sequencing of the
prion gene revealed susceptible haplotypes to scrapie disease in Lebanese goats and one novel haplotype
at amino-acid positions of 136, 154, and 171, corresponding respectively to alanine, histidine, and
arginine. Such polymorphism in MHC class II DRB and in prion genes of experimental goats could be
used in selection for better immunity and resistance to scrapie.
The optimized standard PCR for Growth Hormone (GH) gene gave better yield of the two amplicons
compared to the optimized multiplex system. The HaeIII restriction enzyme succeeded in uncovering the
high polymorphism in the Growth Hormone gene of the local goats. The banding pattern of the restricted
amplicons of GH-exon 4 of local goats was previously correlated with productive performance. The
three different banding patterns of restricted amplicons of exon 2 and 3 of GH-gene were not reported
before, requiring their future correlation with production. The optimized PCR for αS1-casein gene of
local goats revealed two different positions of the amplicons, a result that was documented previously.
The XmnI restriction enzyme of Casein gene amplicon resulted in four different polymorphism,
reflecting the high variability in the αS1-casein gene sequence. The genotypes of the casein gene
polymorphism were defined, allowing for future selection of Lebanese goats that produce low or high
levels of αS1-casein, in accordance with the various degrees of casein-digestibility in different human
populations that consume goat milk.
Publications & Communications - ‫المنشورات والمساهمات في المؤتمرات‬
1. E.K. Barbour, M. Saadé, F. Sleiman, H. Shaib. Selection for immunity improvement and
prion scrapie control in goats based on respective sperm-genetic markers polymorphism
and nucleotide sequences. CIOSTA XXXIV-Vienna; June, 29-July, 1 2011.
2. M. Saadé, E.K. Barbour, F. Sleiman, S. Hamadeh, H. Shaib Selection for production
improvement in goats based on sperm-genetic markers polymorphism. CIOSTA
XXXIV-Vienna; June, 29-July, 1 2011.
3. Elie K. Barbour, Maya F. Saade, Fawwak T. Sleiman, Shady K. Hamadeh, Youssef
Mouneimne, Zeina Kassaifi, Ghazi Kayali, Lina S. Jaber and Houssam A. Shaib.
Quantitative optimization of Saanen goat-sperm genes amplification: evaluation of
standardized protocols applicability in genetically-uncharacterized rural goats. Tropical
Animal Health and Production (UK). In review.
Abstract - ‫موجز عن نتائج البحث‬
The purpose of this research is to optimize quantitatively the amplification of specific sperm
genes in reference genomically-characterized Saanen goat and to evaluate the standardized
protocols applicability on sperms of uncharacterized genome of rural goats for inclusion in
future selection programs. The optimization of the protocols in Saanen sperms included three
production genes (Growth Hormone (GH)-exons 2, 3, and 4, αS1-Casein (CSN1S1), and αLactalbumin), and two health genes (MHC Class II DRB and Prion (PrP)). The optimization
was based on varying the primers concentrations, and the inclusion of a PCR cosolvent
(TritonX).
The impact of the studied variables on statistically significant increase in the yield of amplicons
was noticed in 4 out of 5 (80%) optimized protocols, namely in those related to GH, CSN1S1,
α-Lactalbumin, and PrP genes (P<0.05). There was no significant difference in the yield of
amplicons related to MHC Class II DRB gene, regardless of the variables used (P>0.05).
The applicability of the optimized protocols of Saanen-sperm genes on amplification of
uncharacterized rural goat sperms revealed a 100% success in tested individuals for
amplification of GH, CSN1S1, α-Lactalbumin, MHC Class II DRB genes, and a 75% success
for the PrP gene.
Selection for immunity improvement and prion scrapie control in goats, based on a respective
sperm-genetic markers polymorphism and nucleotide sequences, is concluded in Saanen and
Local breeds. Standardization of the amplification of the second exon of the caprine MHC
classII DRB gene, followed by PstI and TaqI restriction endonucleases reactions, confirmed the
polymorphism in this marker that could correlate with different levels of immunity during
selection of goats.
Selection for production improvement in goats, based on specific sperm-genetic markers
polymorphism, is concluded. The amplified genetic markers that were subjected to restriction
endonuclease reactions included genes of the Growth Hormone and αS1-casein. Amplification
standardization was based on reading the concentration of the amplicons resulting from many
trials, while polymorphism was detected by images of the documented banded fragments
resulting from the endonucleases activities.
The significant success in applicability of the Saanen-quantitatively optimized protocols to
other uncharacterized genome of rural goats, allows for their inclusion in future selection,
targeting the sustainability of this farming system, and the improvement of the farmers
livelihood. In addition, the amplification of the prion gene of goat sperms is standardized,
revealing nucleotide sequences that could be used as markers during selection for elimination of
susceptibility to scrapie disease. The significance of the polymorphism detected in genes of the
Growth Hormone and that of αS1-casein in correlation to production during the selection
process is discussed.