1.
Identification Of Molecular Markers In Bmp15 Gene Of Different Pakistan Sheep And Goat Breeds
by Ahmad Nawaz | Prof.Dr.Masroor Elahi Babar | Prof. Dr | Prof. Dr. Khalid Javed.
Material type: ; Format:
print
; Nature of contents: Publisher: 2011Dissertation note: Genetics of prolificacy in sheep and goat emphasize the importance of main genes which have been made known to affect litter size and rate of ovulation through various mechanisms. Natural mutations in prolific sheep and goat breeds have shown that the transforming growth factor beta (TGF-?) super family ligands such as bone morphogenetic protein 15 is crucial for ovulation and as well as for increasing litter size. Keeping in view the importance of prolificacy in sheep and goat, a research project was planed to identify the polymorphism, its association with fecundity and uncovering the nucleotide picture of BMP15 fecundity gene in sheep and goat breeds of Pakistan. In the research finding, various polymorphism, insertion and deletion of nucleotides in goat and sheep breeds of Pakistan were identified and associated with fecundity and secondly, some novel polymorphism in Pakistani goat and sheep breeds were identified which are different from the goat and sheep breeds of the world. This is the first report of the whole nucleotide of BMP15 gene in the sheep. A lot of work has been reported on these genes but total nucleotide picture in the sheep is not reported. Sequences of Bmp15 gene from sheep and goat breeds of Pakistan has been submitted to the NCBI GenBank database libraries,USA under accession numbers JN655669, JN655670, JN655671 and JN655672. It will result in fast vertical expansion of small ruminants to increase the mutton production and uplift the socio economic condition of small ruminant's farmers in the country.
Availability: Items available for loan: UVAS Library [Call number: 1421,T] (1).
2.
Genetic Characterization Of Pakistani Buffalo Breeds By Mitochondrial D-Loop And Microsatellite Analyses
by Tanveer Hussain | Prof.Dr.Masroor Elahi Babar | Dr. Khalid Javed | Prof. Dr. Irshad Hussain.
Material type: ; Format:
print
Publisher: 2008Dissertation note: Pakistan has various dairy breeds of buffalo and cattle, but the genetic data of different buffalo breeds like Nih, Ravi, Nihi-Ravi, Kundi and Azakheli is lacking which need to be established for their genetic characterization. Blood samples of unrelated true representatives of all breeds were collected from their respective home tracts i.e Nih Ravi (LPRI Bahadarnagar, Okara, BRI Pattoki, Rakh Dera Chahi, Lahore); Nih (Pakpatan,
Minchnabad, Arifwala, Hasilpur); Ravi (Kamahia, Tandlianwala); Kundi (Tandojam, Tando Muhammad Khan, Dadu) and Azakheli (Directorate of Livestock Research & Development Station Surezai, Peshawar and Matta, Swat). DNA was extracted with the use of standard protocol and amplification of the mitochondrial D-loop region was done with specific primers in Molecular Cytogenetics and Genomics Laboratory in the department of Livestock Production. Sequencing of amplified portion of mt DNA D-loop was done. Sequences were analyzed with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNPs) were identified and comparison of 52 mitochondrial DNA haplotypes of all buffalo breeds was done. Genetic distance and identity between five buffalo breeds were calculated and phylogenetic tree was constructed using BioEdit and MEGA 4.1 softwares showing the relationships between different haplotypes. Domestication events were also observed through network analysis. For further confirmation of the genetic structure of buffalo breeds 8 dye labeled microsatehhite markers (recommended by ISAG) were used and genotyping was done. Results were analyzed with the help of different softwares. Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Neis genetic identity and genetic distance/ diversity was calculated. This work provided the genetic data which is very helpful for determining the genetic diversity of buffalo population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future.
Availability: Items available for loan: UVAS Library [Call number: 1114,T] (1).
3.
Identidiation Of Genetic Susceptiblity Of Myopic Loci In Families From Punjab
by Maria Fareed Siddique | Prof.Dr.Masroor Elahi Babar | Dr. Sehrish Firyal | Prof. Dr. Abu.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Myopia, or nearsightedness, is a condition in which the eye cannot focus on distant objects and sometimes closer ones too. In past different authors reported different loci responsible for myopia. They used specifically synthesized markers for different loci and after conducting linkage analysis through genotyping the myopic families were found to be linked for those loci, whereas, in some studies the cause of myopia was environmental. Till now, linkage studies have identified at least 18 possible loci in 15 different chromosomes associated with myopia, although some of these remain to be confirmed. In past, no study was done in Pakistan on myopic families for finding responsible myopic locus in this regard. So, more conclusive and well-designed studies on family pedigrees of individuals with high myopia were needed to be conducted in Pakistan by using genetic markers associated with myopia.
In this study, a panel of microsatellite markers was developed. Blood samples were taken from six myopic families. DNA was extracted. PCR was performed for amplification of these I microsatellite markers on 34 samples belonging to 6 families. Genotyping analysis was performed for the PCR products of microsatellite markers. These results were studied by constructing and analyzing haplotypes on the basis of PAGE gel bands. Heterozygosity, homozygosity, polymorphism with all microsatellites markers, specific for two loci were checked.
One family MYO-4 was found to be potentially linked with markers for the locus MYP-18. Another family MYO-5 showed potential linkage for the locus 2q37.2. Remaining four families (MYO-l, MYO-2, MYO-3 and MYO-6) were totally unlinked with all the markers (D14S984, D14S63, D14S999, D2S2202, D2S2968 and D2S338 for both loci demonstrating genetic insusceptibility of myopic loci in developing myopia and thus suggesting the complex genetic variability of myopia.
This study will serve as the pioneering database for further research on identifying the genetic heterogenic complexity of myopia. Results of this study lead to development of a panel of microsatellite markers which can be used for linkage studies of more myopic families in Pakistan. This study opens the door for new geneticists as the results can also be helpful in carrying out genetic counseling for the myopic persons who are going to be married and specifically for those who have dominant inheritance. This was a preliminary study on myopic patients in Pakistan and data produced during this study will be helpful for drawing and determining genetic inheritance of expected babies with affected parents and siblings. Moreover this study can become the basis for further research investigations on myopics in Pakistan.
CONCLUSION
This was a pioneering study to develop panel of microsatellite markers for conducting linkage analysis and genetic characterization of myopic patients in Pakistan. As a result of this successful study a reliable, efficient and very informative panel of microsatellite markers was successfully developed which was capable to interpret individual diseased allelic identity, to be used for conducting linkage analysis through genotyping of myopics in Pakistan. This study will serve as the database for further research on identifying the genetic heterogenic complexity of myopia and also these successful results can be further analyzed in future on more myopics from different areas of Pakistan. This work provokes the need for further research purposes in identifying the genes influencing myopia that could help develop targeted treatments for children who are genetically predisposed to developing myopia.
Availability: Items available for loan: UVAS Library [Call number: 1177,T] (1).
4.
Dna Fingerprinting Of Pakistani Buffalo Breeds (Nili-Ravi, Kundi) Using Microsatellite And Cytochrome B Gene
by Rashid Saif | Prof.Dr.Masroor Elahi Babar | Mr. Asif | Prof. Dr. Abu Saeed Hashmi.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Customarily, classification of breed was based on phenotypic traits. In some cases, recent genetic studies have found differences in the structure proposed. In buffalo ,one has to bear in mind that morphological changes were not the result of adaptation to the environment, but have a social significance thus may not be indicative of the genetic relationship. In recent years microsatellites have proved to be very useful for the determination of genetic relationship among population. Comparative studies beiween microsatellite and protein markers have highlighted the advantages of the former.
The water buffalo (Bubalus bubalis) holds tremendous potential in livestock sector in many Asian countries, particularly in Pakistan but the genetic data of different buffalo breeds like Nili-Ravi and Kundi is lacking, which need to be established for their genetic identification. Blood samples of unrelated true representative of both breeds (Nili-Ravi and Kundi) were collected from different government livestock farms in Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the mitochondrial Cytb gene and microsatellite was done with especially designed primers in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore. Cytochrome b gene markers have been proved as an efficient and powerful tool for breed characterization and species identification of buffaloes. Several panels of microsatellite markers have also been reported for this purpose. In this study, a panel of nine microsatellite markers has highly Polymorphism Information Content (PlC) were selected, Specific primers were designed for these microsatellite and Cytb gene partial amplification using primer3 software. Then primers were optimized for successful amplification with minimum reagent concentration. PCRs were performed for amplification of these microsatellite and Cytb markers on each sample, Genotyping and sequencing was conducted on all amplicons to find out the different SNP to design haplotypes with the help of bioinformatics software e.g. Blast 2sequence and Chrornas Lite, Further statistical analysis was done by the help of some other software e.g. Popgene version 1.31, Power Stat., Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance! diversity was calculated. The results obtained from this study can contribute to the establishment of routine DNA typing services, beneficial for the buffalo industry as well as in animal forensics for litigation and expedite the police investigation services in Pakistan, which will also be useful for breed characterization and phylogenetic study of aforementioned breeds of buffalo.
Availability: Items available for loan: UVAS Library [Call number: 1183,T] (1).
5.
Molecular Diversity Analysis Of Sheep And Goat Breeds Of Pakistan Using Microsatellites.
by Misbah Shaheen | Prof.Dr.Masroor Elahi Babar | Mr. Tanveer Hussain | Prof. Dr. Azhar.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Pakistan is rich in Animal Genetics Resource (AnGR) and has various breeds of sheep and goat but the genetic data in these different breeds is lacking which needs to be established for their genetic identification. The advent of molecular techniques has led to an increase in the studies that focus on the genetic characterization of domestic breeds using genetic markers. Due to their reliability and availability, the microsatellites have become preferred method for the genome mapping. Microsatellites or STRs are the 1-6 nucleotide tandem repeats present in both coding and non coding regions of both prokaryotes and eukaryotes. Microsatellites are powerful tools in genome mapping, forensic DNA studies, paternity testing, population genetics and conservation! management of biological resources. The present study was conducted on the molecular diversity analysis of sheep and goat breeds of Pakistan using FAQ recommended unlabelled microsatellites. Blood samples of unrelated true representative animals of two sheep and goat breeds were selected from their breeding tracts and different Government Livestock Farms throughout the country. DNA was extracted with the standard protocol and amplification of DNA was done with a set of 16 microsatellite markers in Molecular Cytogenetics and Genomics Laboratory in the Institute of Biochemistry and Biotechnology. The products of touch-down PCR were examined on non denaturing Polyacrylamide Gel Electrophoresis (PAGE). Genotyping results were analyzed through the sofiware POPGENE version 3.3 for calculating the number of alleles, expected and observed heterozygosity, homozygosity, Polymorphic Information Content (PlC).
Average observed heterozygosity, average observed homozygosity, observed and effective number of alleles for all loci and populations were 0.8394, 0.1606, 3.6875 and 2.8693 respectively. Almost all of the microsatellite markers showed significant variations in both breeds of sheep and goat. Genotyping results of microsatellite markers were clearly different for four different breeds showing a distinct genetic distance between sheep and goat breed's.
This work provided the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in future according to FAO global Farm Anithal Genetic resource data. Moreover this study can become the basis for further research investigations in sheep and goat in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1200,T] (1).
6.
Linkage Analysis Of Myp12 And Myp14 In Families From Lahore
by Maryam Zahra | Prof.Dr.Masroor Elahi Babar | Dr. Ali Raza Awan | Prof. Dr.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Myopia is one of the most common refractive errors of the eye worldwide that can effect clarity of vision, limit occupational choices and contribute to increased risk to vision threatening conditions. Six families of different casts were enrolled from Lahore (Punjab). Total of six autosomal dominant families were screened for linkage to the known nonsyndromic autosomal dominant and QTL myopia locus, MYP12 and MYP14 respectively. 5mL blood sample were collected aseptically in a 5Oml falcon tubes containing EDTA. DNA extraction was done by inorganic method. Three markers for each locus were selected from literature and redesigned by using 'primer 3' software. These markers were optimized for their annealing temperature and specific concentration of PCR ingredients by gradient PCR. After that all of the markers were amplified separately on genomic DNA samples of each family. PCR products of each of the marker were run on 1.2 % agarose gel along with 50 base pair ladder to visualize the bands of amplified products at 110 volts for 30 minutes. Linkage analyses were carried out by genotyping through PAGE unit of Major Science, model no. MV-2ODSYS. PCR products were run on Polyacrylamide Gel Electrophoresis (PAGE) to examine amplified bands of microsatellites. A standard 5Obp DNA ladder was run along with the sample PCR products as a reference. By reading the alleles appeared on gel haplotypes were constructed for each member of these families. The overall results of this study did not show evidence for linkage of myopia in thee families to the selected loci MYP12 and MYP14 on chromosomes 2 and 1 respectively. It might be possible any other identified locus or any new locus involved in this population of Pakistan. The findings represented here do not represent the conclusion of this study but do provide ongoing data for further investigation into the exact gentic causes of mypia in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 1216,T] (1).
7.
Identification Of Single Nucleotide Polymorphisms In Olri Gene And Its Association With Milk Composition In Cattle Breeds in Pakistan
by Nusrat Majeed | Prof.Dr.Masroor Elahi Babar | Dr. Ali Raza Awan | Prof. Dr. Abu.
Material type: ; Format:
print
Publisher: 2011Dissertation note: Pakistan being agriculture based country has a great potential in livestock sector. it plays
an important role in the economy of the country. Milk is a balanced diet because it
contains all the essential nutrients like carbohydrates; fats, proteins, vitamins, minerals
and enzymes required for health. In these milk nutrients fat is second most important
component of milk. Primary component of milk fat is triglycerides (triacylglycerols or
TAG) a typical storage form of lipids. OLRI is the major protein that binds, internalizes
and degrades oxidized low density lipoprotein. Ol.Rl as a protein important for oLDL
metabol ism may contribute to these effects. Oxidized fat inhibits the expression of
lipoprotein lipase and fatty acid transporter genes that causes a reduced uptake of fatty
acids into mammary glands. As a result the concentration of triacylglycerols in milk is
reduced. OLRl as a protein important for oLOL metabolism may contribute to these
effects. OLR J identified as a functional and positional candidate gene product for milk fat
percentage and milk fat yield. Polymorphism in Ol.Rl gene increases the milk fat
percentage. If polymorphism in Ol.Rl gene is found in Pakistani Sahiwal and Ohanni
cattle breeds these were help to screen the animals at younger age and also be used to
characterize the different cattle breeds as a milk fat production marker. Unrelated animals
were selected for this study. Blood samples were collected from different Go\'1. livestock
farms/experimental stations. DNA was extracted by organic method. Primers were
designed by using Primer3 software. After DNA amplification by polymerase chain
reaction. peR product was sequenced. Sequencing results of the full length OLR I gene
were analyzed by alignment of sequences with the help of Blast2sequence software.
Novel S Ps were identified. Total of twenty one SNPs were identified in the samples of
all breeds that were confirmed at population level by sequencing of each sample. Then
total twenty one SNPs were found in all breeds. Most of SNPs were found in intronic
regions. In Sahiwal only 5 SNPs are exonic and one of them is common in both Sahiwal
and Red Sindhi breeds. Out of total five exonic SNPs, only two were found synonymous
and rest of the three exonic SNPs were found to be non synonymous. while all other
SNPs are intronic in nature. An intronic SNP is also common between Sahiwal and
Dhanni breed. In Red Sindhi a SNP was observed at 3'UTR. Results were analyzed by.
using the statistical software POPgene32. The mean value of expected heterozygosity that
is 0.3595 is greater than the mean of observed heterozygosity that is 0.0300 that shows
low variabil ity in breeds. The mean value for Shannon's Information index (1*) is 0.5421.
The aim of research is to identify gene that underlie the genetic variation in bovine milk.
fat composition.
Availability: Items available for loan: UVAS Library [Call number: 1396,T] (1).
