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1. Sequence Analysis Of Comt Gene As Suceptibility Factor For Aggression In Domestic And Wild Cats

by Maham Nawaz (2011-VA-455) | Dr. Asif Nadeem | Dr. Saadat Ali | Dr. Amjad Riaz.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Behavior that is directed to injure living beings and damage their neuroconductual processes without any incitement, represents the aggressive behavior. COMT gene is of much importance in determining violent act in both animal and human.The present study is designed to molecular characterize the gene with following objective: to screen out polymorphism (SNPs) in exonic region of COMT gene in cats and tiger and to associate the identified polymorphism in cats and tiger. Aggression questionnaire was filled by honors of all domestic and wild cats included in our study.Blood sample of 5 Stray cat, 5 Persian cat and 5 Siamese cat and 5 Bengal tigers were collected from Lahore Zoo, UVAS PET Centre, Private Pet Clinics and Safari Zoo Lahore for SNP analysis. DNA were extracted from blood by organic method, 5 sets of primers were designed by primer 3 software for the amplification of the COMT genes. The amplified PCR products were precipitated and sequenced bi-directionally on ABI 3130XL Genetic analyzer, for the identification of SNPs.Alignment of sequences were done with the help of blast2 sequences. For sequence data analysis, Bioedit and Clustal W were used to complete the study.Results of sequencing were analyzed using BioEdit software. Sequence alignment tool like Clustal W was used for SNPs identification. 3 intronic and 1 exonic SNPs were observed and confirmed by Clustal W. Exonic SNP was linked to aggression. However, intronic SNPs were not found to be associated with self-reported aggression. SNP observed in exon 2 is reported to be involved in psychiatric and depressive disorders.Our study highlighted the role of COMTgene polymorphisms in aggression in animals (Cats and tiger). Different breeding Policies and Pet plains are now working and we can screen out the susceptibility of aggression in cats and tiger. Availability: Items available for loan: UVAS Library [Call number: 2901-T] (1).

2. Development Of Novel MTDNA Metbarcodes For Species Differentiation Of Class Pisces

by Hira (2010-VA-476) | Dr. Muhammad Imran | Dr. Saadat Ali | Dr. Amjad Riaz.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: The Folmer COI mtDNA universal primers that are considered standard for DNA barcoding of life contain so many mismatches against the target sequences of vertebrate origin that they often end in failure to amplify many of vertebrate DNA extractions. This discrepancy favors for the selection and designing of new metabarcode primers that can be used to identify all individuals of vertebrates or at least all individuals represented in a class of Vertebrata such as Class Pisces. The current study embarks on such an endeavor. In this study development of new mtDNA metabarcode (16SrRNA) that can be used as universal primers to amplify almost all species of Class Aves for different forensic and molecular biodiversity analyses. Fins/tissue samples were collected from Class Pisces (one specimen from every order reported to be present in Pakistan). DNA was extracted from the collected specimens through standard organic method, qualified and quantified and then PCR-amplified using novel universal primers selected from aligned mtDNA sequences originating from all Pisces mitochondrial DNA genomes submitted to different online sequence databases such as NCBI nucleotide database. The sensitivity of PCR also be assessed using a range of DNA concentrations. The amplified products were sequenced on ABI Genetic Analyzer following Sanger’s dideoxy method of sequencing. The correctness of obtained mtDNA sequences were examined visually in Chromas Lite 2.1 software and then alignment of these sequences were performed against highly similar DNA sequences in NCBI nucleotide databases using BLAST in order to identify origin of unknown mtDNA sequences. With the help of sequencing and phylogenetic studies specificity of the universal primer set confirmed and presented as a novel metabarcode (16SrRNA) for species level identification of large number of Piscean species In summary, we present universal method for species classification of Pisces using a targeted parallel sequencing approach. Both sequencing and phylogenetic studies experiments confirm specificity of universal primer set. Although promising results were obtained with current settings, rapid improvement of bench top instruments will further develop method with less hands-on, fewer sequencing errors and lower detection limit. So, in future, this barcode can be used for species identification in various fields of study such as meat adulteration, illegal trade, food mislabeling and molecular estimation of biodiversity. Availability: Items available for loan: UVAS Library [Call number: 2899-T] (1).



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