1.
Molecular Investigation Of Mental Retardation Locus (Mrti)/Gene Prss12 By Linkage Analysis
by Zafar Ali | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Mental retardation (MR) is a condition in which a person having an intelligence quotient (IQ) lowers than 70. It is also associated with a deficit in adaptive behavior such as communication and daily living skills. Mental retardation is either non-syndromic or syndromic. It is one of the most common genetic disorders and it affects about 1-3% of the human population, with a ratio of males higher than females. The present study was can-ied out to determine the prevalence of families having mental retardation in Pakistani population. In the present study, 7 MR families with three or more affected individuals with MR were enrolled. Family history was taken and pedigree was made personally by visiting the families. The blood samples were collected from the enrolled families. Then DNA was extracted from the blood samples collected from these families by standard inorganic protocol. After isolation of DNA from blood samples, 3 STR markers (D4S191, D4S2392 and D4S3024) in vicinity of mental retardation (MR) locus (MRT1)/gene PRSS12 were amplified on each sample of each family by PCR. The amplified PCR product was first checked on agarose gel and then genotyping analysis (linkage analysis) was performed on non denaturing polyacrylamide gel (PAGE). After polyacrylamide gel electrophoresis, picture of the gel was taken and alleles were read manually with larger allele donated by 2 and smaller by 1. After that haplotype was constructed to determined the pattern of inheritance among the affected and normal individuals of each family under study and also to determined that a family was linked or unlinked to mental retardation locus (MRTI)/gene PRSS12. None of the family was linked to mental retardation locus/gene PRSS12. The families which remain unlinked to the reported loci during screening signifies extreme genetic heterogeneity of MR which is not surprising because about 50% of human protein coding genes are expressed in the brain and it provides an excellent resource material for mapping of the new genes which will shed light on the complex pathways involved in the development of learning and memory in those population. The pedigree of each family in the present study showed that most of the marriages are cousin marriages; therefore this study may play a role in creating awareness about the effect of cousin marriages that is the first step towards decreasing socio-economic burden of the country by genetic counseling and also to prevent mental retardation in Pakistan due to inbreeding. Mental retardation locus (MRT1)/gene PRSS12 was studied for linkage analysis in seven families from different areas of District Swat and Peshawar of Khyber Pakhtunkhwa province of Pakistan. None Out of seven families was linked to mental retardation locus (MRT1 )/gene PRSS 12. All the seven families remain unlinked to this locus. It is concluded that Mental retardation is a complex genetic disorder and needs further studies to identify the already known locus or to explore novel loci through genome wide scan responsible for mental retardation in these population. This will provide opportunities of genetic counseling to these populations and will ultimately result in prevention of mental retardation in Pakistani population.
Availability: Items available for loan: UVAS Library [Call number: 1171,T] (1).
2.
The Study Of Gene Gjb2/Dfnb1 Causing Deafness In Humans By Linkage Analysis From District Peshawar
by Noor Badshah | Prof.Dr.Masroor Elahi Babar | Dr. Aftab | Mr. Muhammad Asif.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Hearing impairment is the partial or complete inability to hear that leads to compromise the development of normal language skills. Among all the sensory impairments in humans, hearing impairment is the most common.
It is estimated that at least 50% of the cases are due to genetic factors. Hereditary hearing loss may be syndromic or non-syndromic; about 30% of deafness cases are syndromic, while 70% is non-syndromic. It is estimated that the prevalence of profound bilateral hearing loss is 1.6 per 1000 in Pakistan and 70% of hearing loss arises in consanguineous families. The main pattern of inheritance of deafness in Pakistani population is autosomal recessive and to date more than 145 loci and 26 genes have been identified for non-syndromic recessive deafness.
More than 400 disorders associated with hearing loss shows extreme genetic heterogeneity and complexity of the mammalian inner ear. As more genes are identified, the elucidation of the function of the proteins that these genes encode contributes greatly to the understanding of cochlear mechanisms and their role in disease causation.
The gene involved, GJB2, encodes the connexin26 molecule. Connexin26 is a component of gap junctions, the links that allow small molecules to pass from one cell to the next, and this protein is found in several places in the body, including the epithelial supporting cells surrounding the sensory ear cells of the cochlea.The sensory ear cells of the cochlea allow potassium ions to pass through their upper surface during normal reception of sound, and these potassium ions must be recycled through the base of the ear cells and the supporting cells and fibrocytes back into the high-potassium endolymph that bathes the tops of the ear cells.
The aim of this study was Linkage analysis for DFNB1 locus involved in causing hereditary deafness in families from Khyber Pukhtunkhwa. A total of 10 families were enrolled from different areas of Khyber Pukhtunkhwa province. I have studied 8 families of these 10 (i.e.) family no. 2, 3, 4, 5, 6, 8, 9 and 10. The families have at least three affected individuals. All the families showed recessive mode of inheritance. For linkage analysis studies for DFNB1 locus, three STR markers D13S175, D13S292, and D13S787 were genotyped using Polyacrylamide gel electrophoresis (PAGE) and haplotypes were constructed to determined, linkage with DFNB1 locus. From a total of 8 families, a single family was linked to DFNB1 locus. The DFNB1 locus is the first non-syndromic deafness locus mapped to chromosome 13q12.
Availability: Items available for loan: UVAS Library [Call number: 1191,T] (1).
3.
Genetic Diversity And Differentiation Of Domestic Buffalo Of Pakistan Through Sky And Zfy Genes Of Y Chromosome
by Muhammad Mudassar Manzoor | Mr. Taneer Hussain | Mr. Muhammad Asif | Prof. Dr. Abu.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2011Dissertation note: Livestock sector plays a vital role in the economy of Pakistan. Main contribution of milk comes from buffaloes and cows. Water buffalo (Bubalusbubalis) is one of the major elements of livestock in the country and possess great importance for economy in the form of milk and meat productions. Nili, Ravi,Nili-Ravi, Kundi and Azakheli are major breeds of water buffalo to be found in different areas of Pakistan. Conventional classification of breeds 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 Y chromosomal genes have proved to be very useful for the determination of genetic relationship among population. Comparative studies have highlighted the advantages of the SRY and ZFY genes of Y chromosome.These genes have been considered as competent and powerful tool for the purpose of breed characterization and species identification of buffaloes. In livestock sector, water buffalo (Bubalusbubalis) has shown great prospective in numerous Asian countries including Pakistan. Unfortunately, there is lack of genetic data of different buffalo breeds like Nili-ravi and Kundi which needs to be established for their genetic identification.Blood samples from true representative animals of each of the two buffalo breeds (Nili-Ravi and Kundi ) were collected from different Government livestock farms and their respective home tractsin Punjab and Sindh respectively. DNA was extracted by inorganic method and amplification of the SRY and ZFY(exon 5) genes of Y chromosome was done with especially designed primers using Primer3 software in Molecular Biology and Genomics Laboratory at Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore.Specific primers are designed for these genes amplification. Then primers were optimized for successful amplification with minimum reagent concentration. PCR was performed for amplification of SRY and ZFY(exon 5)genes on each sample. Sequencing was conducted on amplicons to find out the different single nucleotide polymorphism (SNP) to make haplotypes with the help of bioinformatics software like Blast 2sequence and Neighbour Joining phylogenetic tree was constructed by using MEGA version 5 (Tamuraet al. 2011). The results obtained from this study now can contribute to the establishment of routine DNA typing service to the advantages of the buffalo in livestock industry.
Availability: Items available for loan: UVAS Library [Call number: 1378,T] (1).
4.
Molecular Characterization Of Local Isolation Of Staphylococcus Aureus On The Bsis Of 16S Rrna From Poulry And Their Transmission to Humans
by Muhammad Rizwan Ashraf | Mr. Muhammad Asif | Dr. Aby Saeed | Mr. Tanveer Hussain.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Staphylococcus aureus is a widely distributed throughout the world and makes up the normal bacterial flora of skin and mucous membranes of man and animals. It is involved in suppurative wound infections in man and animals. Poultry industry has also been effected by S. aureus and causing great economic and health problems. The focus of the microbiology is to correctly identify S. aureus for the treatment of the animals. Molecular biology and biotechnology is proving a helping hand in the accurate identification of microorganisms through sequence analysis of 16S rRNA gene. The aim of this study was the molecular characterization of S. aureus from poultry and poultry farm workers through 16S rRNA analysis. Bacteria were collected from poultry and poultry farm human workers. All the samples were cultured and tested biochemically. In addition, peR amplification of 16S rRNA was performed in order to sequence the gene and further analyses through bioinforrnatics tools were performed. The aim of the study was the molecular characterization of S. aureus in poultry and humans through 16S rRNA sequencing, finding the phylogenetic relationships among S. aureus isolates and detection of zoonoses between poultry and human. 16S rRNA gene was amplified with peR primers and the sequence was compared with NeBI database reported S. aureus sequences. Resemblance was found between human and chicken isolates. Phylogenetic analyses were performed by using MEGA5 so ftware that also showed phylogenetic relationship among them.
Availability: Items available for loan: UVAS Library [Call number: 1393,T] (1).
5.
Designing Of Oligo Pool All For The Selection Of Superior Dairy Animals In Pakistan
by Kamran Abbas | Prof. Dr. Masroor Ellahi Babar | Dr. Aftab | Mr. Muhammad Asif.
Material type: ; Format:
print
Publisher: 2011Dissertation note: Livestock has an important role in an agriculture based country like Pakistan with a large number of dairy animals. However the average daily milk yield of dairy animals is very low. There is need to improve milk yield by the selection of superior dairy animals using latest genomic selection procedures. Selection of superior animals on the basis of genetic markers has a tremendous potential for breed improvement across the globe. Substantial advances have been made over the past decades through the application of molecular genetics used in industry programs for several decades and is growing, the extent of use has not lived up to initial expectations. Most applications to date have been integrated in existing programs on temporary basis.
Among various molecular markers the Single Nucleotide Polymorphism (SNP) is one of the major genetic marker used worldwide. Through SNP genotyping selection of phenotypic superior animals can be done. There are many techniques used for SNP genotyping but the most advanced technique is Veracode GoldenGate Assay by Illumina. Illumina's VeraCode technology with the BeadXpress (BX) Reader is ideal for high-throughput small to mid-scale genotyping studies and SNP validation. BX leverages the power of digital holographic codes and the robust GoldenGate Genotyping Assay to provide a detection method for multiplex assays requiring high precision, accuracy, and speed. A custom assay of 48, 96, 144,192 and up to 384-SNPs OPA (Oligos Pool All) is designed using Illumina's Assay Design Tool and manufactured by Illumina.
As a first step for designing of Veracode GoldenGate Assay the development of Oligo Pool All (OPA) is necessary. The OPA was designed by using the genes for milk production, growth, fertility, health and other performance traits. The SNP's in these genes was searched from different gene banks and after proper arrangement the files were sent to Illumina for scoring. After scoring the OPA was finalized for the Veracode GoldenGate Assay for the selection of superior dairy animals in the country using the highly robust BeadXpress technology. The development of OPA for the selection of superior dairy animals was done for the very first of its kind based on modern technology, Veracode GoldenGate Assay in Pakistan. This will greatly help the livestock and dairy development departments, livestock owners, breeders, forensic agencies and researchers to use this unique panel of molecular markers for the selection of superior animals on the basis of marker assisted selection.
Availability: Items available for loan: UVAS Library [Call number: 1528,T] (1).
