UVAS Library

Your cart is empty.

Your search returned 2 results. Subscribe to this search

Not what you expected? Check for suggestions
Unhighlight Highlight
|
1.
No cover image available
Development Of Novel Mtdna Metabarcodes For The Species Differentiation Of Class Amphibia

by Rehmatullah (2011-VA-365) | Dr. Muhammad Imran | Dr. M. Yasir Zahoor | Dr. Amjad Riaz.

Material type: book Book 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 Amphibia. 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 Amphibia for different forensic and molecular biodiversity analyses. Tissue samples were collected from order Urodela of Class Amphibia (Toads , Bull frog and skittering frogs sample were collected from Punjab, Pakistan). DNA was extracted from the collected specimens through standard organic method, qualified and quantified and then PCRamplified using novel universal primers selected from aligned mtDNA sequences originating from order Urodela 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 Summary 67 presented as a novel metabarcode (16SrRNA) for species level identification of large number of Amphibian species. In summary, we present universal method for species classification of Amphibia 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: 2874-T] (1).

Place hold Add to cart
2.
No cover image available
Mutational Analysis Of Atp7b Gene Responsible For Wilson’s Disease And Its Homology Analysis In Primates And Mouse

by Amama Ghaffar (2011-VA-375) | Dr. M. Yasir Zahoor | Prof. Dr. Huma Arshad Cheema | Dr. M. Imran | Dr. Amjad Riaz.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Copper being an essential element to carry out different cellular processes normally is maintained through proper regulation mechanisms to avoid its accumulation in the body. ATP7B gene that codes for ATP dependent P type ATP7B protein controls the regulation of copper in the body. It is required for the proper delivery of copper to apoceruloplasmin and its excretion through bile in the form of feces. Therefore, mutation occurring in the ATP7B gene can cause excessive cellular copper accumulation which results into Wilson’s disease. Variation in ATP7B gene related to copper transportation leads to Wilson’s disease and transmitted in generation through recessive pattern of inheritance. For this study blood samples of fifteen Wilson’s disease affected patients along with normal individuals of the same family were collected from Children's Hospital & Institute of Child Health, Lahore. DNA was extracted from blood through organic extraction method followed by DNA quantification. Amplification of exons 8, 13, 14 and 18 of ATP7B gene was performed after designing specific primers for these specific regions. Sequencing of amplified products was done through dideoxy chain termination method. A disease causing mutation of ATP7B gene c.3155 C>T; p1052 Proline (CCC) to Leucine (CTC) has been mapped on exon 14 in family with Wilson’s disease. This mutation can be used for genetic testing, prenatal diagnosis and genetic counseling. No mutation was found in exons 8, 13 and 18 which mean that further study needs to be done to find more local mutation(s) that can be used for fast direct genetic testing of Wilson’s disease patients or the carriers with heterozygotic conditions who can develop this disease at any age of their life. Results 87 MUSCLE and Clustal Omega were used for homology analysis of ATP7B gene nucleotide and protein sequences that revealed Gorilla to be closest to human regarding coding sequences, while Clustal Omega output file showed all the species varied highly in their protein structure homologies. Through the prediction of secondary structure homologies it was seen that marmoset was closest to humans. This study helped in providing prenatal diagnosis and genetic screening services in the country. It has facilitated in selecting animal models for further study and research on ATP7B gene and molecular pathogenesis of the Wilson’s disease leading to prevention and cure of disease. Availability: Items available for loan: UVAS Library [Call number: 2892-T] (1).

Place hold Add to cart

Implemented and Maintained by UVAS Library.
For any Suggestions/Query Contact to library or Email:rehana.kousar@uvas.edu.pk Phone:+91 99239068
Website/OPAC best viewed in Mozilla Browser in 1366X768 Resolution.