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51. Comparative Genomic Study of Motor Neuron Disease in Horses and Human

by Shakeela Daud (2011-VA-534) | Dr. Muhammad Wasim | Prof. Dr. Tahir Yaqub | Dr. Abu Saeed Hashmi.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: cd not submitted Availability: Items available for loan: UVAS Library [Call number: 2810-T] (1).

52. 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).

53. Molecular Diagnosis Of Brucella Zoonosis As Blood Transfusion Hazard In Metropolitan Population Of Lahore And Okara

by Amna Azam (2011-Va-3560 | Dr. Wasim Shehzad | Dr. Iahtasham Khan | Dr. Muhammad Imran | Dr. Imran Rashid.

Material type: book Book Publisher: 2017Dissertation note: Brucellosis and Coxiellosis are one of the most spreading zoonotic diseases. They both are facultative, intracellular, Gram negative and involved in bioterrorism and agro-terrorism attacks. Brucella abortus and Brucella melitensisare common among all specie of Brucella. Total two hundred Human Blood transfusion samples were collected from hospitals of Okara and Lahore. Blood was collected in vacutainers (without EDTA). After serum isolation serological test RBPT were performed of all samples. Eighty nine out of two hundred were positive to RBPT with seroprevalence of 44.5% (95% Confidence Interval [CI]: 37.61 – 51.4). DNA extraction was done. The concentration of DNA was analyzed through Nanodrop 2000. Then these samples were subjected to Genus specific Real-time PCR analysis. Forty one out of two hundred were positive to Genus specific Real-time PCR with seroprevalence of 20.5% (95% Confidence Interval [CI]: 14.9 – 26.09). Brucella genus positive samples were subjected to two species specific PCR Brucella abortus and Brucella melitensis respectively. Forty one out of forty one Brucella genuspositive samples were positive to Brucella melitensisand none of the sample was positive to Brucella abortus. These two hundred DNA samples were then subjected to Coxiella specific Real-time PCR analysis and 4 were found positive with seroprevalence of 2% (95% Confidence Interval [CI]: 0.06 – 3.94). Results were recorded in the form of Ctvalue. Results indicate that Real-time PCR is more efficient than RBPT and due to increasing seroprevalence in Blood transfusion samples its screening should be included in normal blood transfusion screening procedure through collaboration with Government to prevent transfusion transmitted infections (TTI’s). Availability: Items available for loan: UVAS Library [Call number: 2873-T] (1).

54. Study Of Arginine Vasopressin (Avp) As A Candidate Gene For Evaluating Silent Estrus Behavior In Nili-Ravi Buffalo

by Muhammad Danish Ahmad (2011-VA-464) | Dr.Maryam Javed | Dr. Asif Nadeem | Dr.Muhammad Zubair Shabir.

Material type: book Book Publisher: 2017Dissertation note: Buffalo is a major contributing animal in livestock and in Pakistan as an agrarian country its economy. Nili- Ravi buffalo also known as “Black Gold of Pakistan” has a high potential of productivity. But its production is often effected by certain reproductive disorders, out of which silent estrus behavior act as a major limiting factor for its production, as it is difficult to proper diagnose of silent estrus it result in low fertility and ultimately yield as low productivity in buffalo. There are a number of reasons involve in silent estrus behavior such as nutrition, environment and genetics. Estrus is a polygenic trait and according to a report about 269 genes are involve in estrus. One of the major effecting gene on estrus is Arginine Vasopressin (AVP), produced by hypothalamus and released by posterior pituitary lobe. Along with a number of role in body it influences the Social behavior neural network, located in limbic system and produce the responses like sexual arousal, partner pairing, mating process and social dominancy. So the AVP accounted as a potential candidate gene for study the silent estrus behavior in Nili-Ravi buffalo. The basic aim to conduct the present study was to identify the Single Nucleotide Polymorphism (SNP) in exonic region of AVP gene and find their association to the silent estrus trait. Fifty samples in blood form of Nili-Ravi Buffalo was taken from B-block research forms UVAS, Ravi campus and Buffalo Research Institute (BRI) Pattoki. DNA was extracted using Phenol Chloroform Iso-amyl alcohol (PCI) based method, then DNA was subjected to PCR amplification, Product was precipitated and sequenced for genetic analysis. To identify the SNPs in obtained sequence the Bioinformatics tools such as BLAST and CHROMAS were applied. The three exonic regions of AVP gene were amplified using site specific primer sets. A total of 6 Summary 58 polymorphic sites were identified, those all were present in exon 1. The bioinformatics analysis using PopGene32 software was performed to analyze the association of identified SNPs to the Silent Estrus Behavior. SNP were analyzed for their effect on trait and one SNP in exon-1 was analyzed for its effect on subjected trait. This genetic characterization of AVP gene may serve as the genetic source for the development of DNA based markers for used in selection of animals with better estrus trait in studies, research and commercial purposes. Availability: Items available for loan: UVAS Library [Call number: 2871-T] (1).

55. A Thesis Submitted In The Partial Fulfillment Of The Requirements For The Degree

by Ayesha Saddiqa (2011-VA-367) | Dr. Asif Nadeem | Dr. Maryam Javed | Dr. Muhammad Zubair Shabbir.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: AIDS epidemic is increasing rapidly in the Eastern Mediterranean Region. Quoting fresh authorized figures collected by the Punjab health department were 97,000 to 125,000 HIV positive people in Pakistan. Number of patients with HIV/AIDS rapidly increased in Punjab. 310 HIV/AIDS cases (35 women and 13 children) have been stated in Punjab in 2014. CCR5 gene is associated HIV infection. Mutation in this gene delayed the progression towards AIDS. In this study blood samples were collected from the laboratory of Punjab Aids Control Program (PACP), primary and secondary health care department, Government of Punjab. Genomic DNA was extracted by using the using FavorPrepTM Blood/Cultured Cell Genomic DNA Extraction Mini Kit. Specific set of primers were designed for the amplification of the targeted gene. The amplified PCR products were precipitated and sequenced for the identification of polymorphisms. Bidirectional sequencing was done for result confirmation. Alignments of sequences were done with the help of NCBI BLAST. Chromas software, Clustal W, UCSC, Bio Edit and SNPedia and Mega 6.0 was used to compile this study. CCR5 32 base pairs allele deletion was found absent in all HIV positive and negative individuals. So, susceptibility of human immuno-deficiency virus type one infection is high in Pakistani population. Genomic comparison was done with non-human primates. Alignment result showed human CCR5 gene homology, 95%, 99%, 94% and 94% with Maccaca mulata (Rhesus Monkey), Pan troglodytes (chimpanzee), Cercocebus atys (sooty mangabey) and Rhinopithecus bieti (black snub-nosed monkey) respectively. So, this homology analysis showed that these non-human primates can be used for development of therapeutic strategies related to human immune deficiency virus. Availability: Items available for loan: UVAS Library [Call number: 2872-T] (1).

56. Mutation Detection Of Cacnb4 Gene Involved In Childhood Absence Epilepsy And Its Comparative Genomics In Mice

by Ayesha Amin (2015-VA-1048) | Dr. Muhammad Wasim | Dr. Sehrish Firyal | Prof. Dr. Aftab Ahmad Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Childhood absence epilepsy (CAE) is due to multiple factors affecting almost 9-12 years children. Depolarization of ion channel activates the channel. CACNB4 gene is affected by epileptic seizures. Disturbance in ion channel can affects different genes as CACNA1G, CACNA1H, CACNA1A, SCN1B, SCN1A,SCN2A and GABA receptor genes. CACNB4 gene has a major role in influencing epilepsy in human. In present study,it is directed to analyze the mutations in epilepsy present in coding region of CACNB4 gene. Collection of blood samples were from Children Hospital, Lahore, Punjab Pakistan from CAE patients of epilepsy. By using standard DNA extraction method, DNA was extracted from samples. Primers were designed for the amplification of exon 3 and 13 of CACNB4 gene. Results were examined after sequencing the samples. BioEdit software was used to study the samples thoroughly. NCBI BLAST was used to align the sequences. It is investigated that the sequences of CAE patients of epilepsy of CACNB4 gene has mutation at position position 258023bp which changes A>G. In protein sequence, the mutation is at position 413 which changes L (Leucine) to L (Leucine). This mutation has no effect because this is a synonymous mutationwhere the codon CUG is changes to CUA, both codes for same amino acid that is leucine, so no effect at all by this change in exon 13. Three mutations are present in the intronic region of exon 13 first, second and third at positions 258184bp A deleted, 258289bp and 258191bp of CACNB4 gene respectively. These all mutations are present in intronic region so has no effect in phenotypes of individual. In conclusion, maximum numbers of samples were needed to observe the effect of mutations and factors that causes epilepsy. This study will now help the researchers to investigate genetic therapies, strategies of genetic counseling and parental diagnosis for the population of Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2924-T] (1).

57. Study On Polymorphism Of Promoter Region Of Bovine Lactoferrin Gene And Its Relation With Mastitis In Nili Ravi Buffalo

by Muhammad Asim (2012-VA-636) | Dr. Sehrish Firyal | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Dairy animals in Pakistan and worldwide are facing most persistent and economically affecting disease mastitis. Only a healthy buffalo can produce good quality milk of physiologically normal composition. Mammary gland inflammation due to mastitis badly affects the quantity and quality of milk and this causes big loss to dairy industry. Even province Punjab bears economic losses of 240 million per annum due to mastitis. Susceptibility and resistance to mastitis is affected by the variation in immunity genes. Among immunity genes Lactoferrin (LF) have important role in immune defense system and perform antibacterial, antiviral and anti-inflammatory function. LF is found in most of body fluids like, milk, blood, tear, saliva, bile and mucous. Polymorphism in promoter region of Lactoferrin gene is associated with mastitis susceptibility and resistance. For screening of the mastitis susceptibility and resistance of dairy buffaloes, LF is a potential candidate gene. The present study was designed for the identification of polymorphism in LF gene associated with mastitis. Blood samples from 20 Nili Ravi buffalos having clinical and subclinical mastitis were selected. Blood samples of normal Nili Ravi buffalos were also collected. DNA was extracted; specific primers for amplification of LF gene were designed with Primer-3 software by using already reported sequence on NCBI. Amplification of LF gene performed by PCR and sequenced the amplicon. A comparative analysis of sequence result was performed by using NCBI BLAST. BioEdit software was used to perform multiple sequence alignment. Comparison analysis of LF gene promoter region shows multiple mutations in in clinical and subclinical as compare to reported sequence (accession no. EF650854) at NCBI. While normal samples sequence results are similar to reference data. These results show LF is a candidate gene for mastitis resistance. Availability: Items available for loan: UVAS Library [Call number: 2923-T] (1).

58. Mutational Analysis Of Cacna1ggene Implicated In Childhood Absence Epilepsy And Its Comparative Genomics In Mice

by Fiza Idrees (2015-VA-803) | Dr. Muhammad Wasim | Dr. Ali Raza Awan | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Childhood absence epilepsy (CAE) is the subtype of Idiopathic generalized epilepsy (IGE). It accounts for 2-8% of patients with epilepsy. The frequency of CAE is more in girls than boys. The percentage of CAE in youngsters is 10-12%. In addition to CACNA1G, many other genes can be the possible cause of CAE. The pattern of inheritance of CAE is polygenic and complex. SNP might be a gain of function mutation in T- channel genes that results in increase T-type calcium channel activity. Ion channel genes and genes for GABA receptors are affected in epilepsy. By using various techniques of molecular genetics mutations have detected in genes of calcium channels (CACNA1H,CACNA1I, CACNA1A, CACNA1G and CACNB4), in genes of sodium channels like (SCN1B, SCN2A, SCN1A ) and genes for GABA receptor (GABRG2 and GABRD ). Gain of function mutation in CACNA1G gene and increased activity of α1G channels are the possible reason for abnormal SWD in absence epilepsy. Aim of this study was to assess acknowledged and/or the novel mutations in CACNA1G gene obtained from local childhood epileptic patients. Blood samples (n=20) were obtained from CAE patients. These samples were collected from children hospital Lahore. Organic method was used to extract DNA from these collected samples. Specific primers were designed for exon 13 and 17 and these exonic regions were amplified using PCR. After PCR, sequencing of PCR products was performed and then sequencing results were analyzed using chromas lite software. It has been observed that CACNA1G gene has two mutations in exon 17. It was noticed that protein sequence was altered and the positions of mutations were 38594bp and at 38635bp 38594bp and at 38635 bp. So SNP was detected and there was a gain of function mutation α1G channel activity. In conclusion, these mutations are responsible for absence seizures in CAE patients. So, it can be concluded that to find out how individuals get affected by these mutations and what factors are involved in causing such mutation, a large scale study should be conducted.In addition, other genes involved in causing epilepsy should also be investigated in local Pakistani Punjab population. As a result of such studies, various diagnostic procedures, strategies for counseling and gene therapies can develop. Availability: Items available for loan: UVAS Library [Call number: 2925-T] (1).

59. Genetic Evolution And Development Of Recombinant Vaccine Against Newcastle Disease For Chicken In Pakistan

by Abdul Wajid (2009-VA-705) | Dr. Muhammad Wasim | Prof. Dr. Tahir Yaqub | Dr. Muhammad Tayyab.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: Newcastle disease (ND) is one of the most contagious diseases of poultry worldwide. The disease is endemic in Pakistan and recurrent outbreaks have been reported in commercial poultry flocks, domestic pet and migratory birds since 1963 an inception of commercial poultry farming in the country. Disease surveillance is necessary to determine the incidence of the disease as well as to identify the etiological agent of the disease status in the region. The analysis of the field data provides a clue for the higher authorities to take steps for the remedy of the devastating outbreak. A virulent form of Newcastle disease virus caused an outbreak in the northern region of Pakistan during the mid of 2011. The virus was identified as a virulent viscerotropic vvNDV and characterize, belonging to the sub genotype VIIi. However, the virus of this genotype is still circulating in the field though the intensity of the strain to succumb the chickens to cause mortality does not exist. The particular thing in this genotype was its susceptibility to other avian species like pheasants, peafowls, ducks turkeys, peacocks, sparrows and parakeets. As this genotype is circulating since 2011 2016 and still spill over in these avian species. Thus for the last five years (2011-16), 3500 healthy, diseased and dead chickens, pheasants, peacocks, turkeys, peafowls, ducks, sparrows, exotic parakeets, rosy-faced parrots, pigeons, and partridges from 750 different locations s were monitored. Samples were collected from the Northern region of the country Punjab, Khyber Pakhtoonkhawa, Azad Kashmir, including Gilgit,Baltitssan and from Southern region, Karachi, Hyderabad , Mirpursakro and other small cities where the poultry farms are located. The samples were collected by the local veterinarians, poultry Assistants and Animal health practitioners who assist during the surveillance program. Samples were also collected from the farmers who brought their birds for inspection in the lab with the details of the 141 farm. Mostly sampling was done where there was reports of NDV outbreak, tissues were collected usually the trachea, spleen and brain, moreover, the pharyngeal and cloacal swabs not only from the infected birds but also from the healthy birds were collected to assess the virus shedding in the flock. Blood samples were also collected (1% of the birds at farm), for serum collection to assess the immune status of the flock using Haemagglutination Inhibition (HI) test and Enzyme linked immunosorbant assay (ELISA). The Survey Form meet the international standard was filled for each farm for recording the information required to find the diagnostic clue as well as the molecular characterization of the isolates. Pool of five pharyngeal swabs were processed after the passage into 9-day old chicken embryonated eggs and confirming the positive HA test and then confirmed by real time PCR (RT-PCR). In addition, sera were tested against NDV by HI and ELISA tests. The targeted samples were sequenced by complete fusion gene and whole genome using 22 pairs of overlapping primers. The observations indicated that the commercial broiler industry is highly susceptible to virulent NDV and confirmed by data available in the laboratory in the survey form. Contrary to that a little is known regarding the maintenance and enzootic trends of vNDV infection level in domestic and wild birds. Poor strategy of the use of vaccines and vaccination as well as the existence of virulent form of NDV in the domestic and pet birds indicate a possibility of the root cause of the ND eruption in the developing countries. A continuous isolation of virulent viruses of the panzootic Newcastle disease virus of sub-genotype VIIi since (2011-2016 from commercial chickens and from various other avian species in the country provide evidence for the existence of epidemiological links intermingling of the strain among them. Therefore, to avoid the huge economical losses in the commercial poultry the second largest industry in Pakistan, their close proximity should be strictly avoided. The mass vaccination of the poultry flocks is in practice in all commercial 142 poultry farms in Pakistan. However, the use and availability of a reliable and standard vaccine, as well as the correct usage of vaccine dose of the live attenuated LaSota vaccine are the key factors to improve their efficacy in the field. Minor outbreaks have been occurring in the field even though a severe outbreak was occurred in 2011-12 collapsed the poultry industry with other pet and wild birds. To minimize the continuity of these minor outbreaks in the field for long time there is a need for more effective vaccine to control the particular genotype of the ND virus. In the present study, DNA vaccine was developed using the SFR-55 NDV strain antigens, in the form of fusion (F) and hemagglutinin-neuroaminidase (HN), namely pcDNA3.1-F and pcDNA3.1-HN. In vitro expression of both genes construct was assessed by reversetranscriptase- PCR (RT-PCR) and western blotting. In the trial an inactivated oil-based emulsion vaccine was prepared using the field strain SFR-55 and compare with the commercial vaccine LaSota strain commonly used by the poultry industry. Birds were divided into six groups, the first two groups were immunized with pcDNA3.1-F and pcDNA3.1-HN alone respectively and third group with was vaccinated with both antigens pcDNA3.1-F+HN. The other two groups were immunized with inactivated (wvSFR-55) and LaSota vaccines as described above, the last group was injected with empty vector as control. The birds were immunized twice at 14 and 21 days of age intramuscularly (DNA vaccine), subcutaneous and eye-drop by inactivated and LaSota vaccines respectively. The birds were challenged with live virulent NDV strain using a dose of 10,000 ELD50/0.1ml per chicken. Results indicate that Inactivated and LaSota vaccines provided high protection (>80%), as compared to pcDNA3.1-F, pcDNA3.1-HN, pcDNA3.1- F+HN gave 70%, 75% and 20% respectively. There was 100% mortality in control chickens. The administration of two vectors expressing F and HN antigens induced high immune response, and provide protection than when used separately. However, the groups immunized with 143 pcDNA3.1-F, pcDNA3.1-F+HN and inactivated vaccine resulted in lower amount of virulent virus shed after challenge when compared to the group immunized with standard LaSota. In summary, the co-administration of both NDV glycoprotein antigens increased protection than use separately. DNA-based vaccine can be used safely to reduce mortality and most importantly lower the risk of virus transmission due to decreased level of virulent virus shedding. Availability: Items available for loan: UVAS Library [Call number: 2910-T] (1).

60. Association Of Tryptophan Hydroxylase 2 Gene Polymorphisms With Risk Of Depression In Homo Sapiens And Equus Caballus

by Sher Sarmad (2015-VA-1062) | Dr. Asif Nadeem | Dr. Maryam Javed | Prof. Dr. TahirYaqub.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: In the biosynthetic pathway for brain serotonin, Tryptophan hydroxylase-2 (TPH2) acts as the rate-limiting enzyme. It is a key element in maintaining adequate serotonin neurotransmission in the central neuron system (CNS). It is broadly discussed as an important candidate gene in multiple psychiatric disorders, especially suicidal behavior and depression. A relationship between TPH2 and major depressive disorder (MDD) has been reported by multiple gene-disease association studies in different populations. Horse can be employed as a valuable candidate for an animal model of depression because it shares environmental factors which are known to cause depression in humans.The hypothesis of this study was that, there is association between TPH2 gene polymorphisms and risk of developing MDD.Blood was collected from each participant.Human (Experimental and Control group) and Horse (Experimental and Control group).DNA wasextracted usingthe standard Phenol Chloroform Isoamyl alcohol (PCI) protocol. Specific set of primers were designed for the amplification of TPH2 gene exons and partial region of the introns. The amplified PCR products was precipitated and sequenced for the identification of variants. For sequence data analysis Chromas Software(2.1) was used along with BLAST available at NCBI and Clustal W program. Multiple alignments were performed for polymorphism identification and association of identified polymorphisms was performed using SPSS.The significant association of the variant rs7305155 with Major Depressive Disorder indicates that TPH2 has a role in disease etiology.Understanding the extent of the role different genes play in MDDmay help in tailoring medication. “Gene-Response to drug” association studies have also shown that patients with certain polymorphisms might respond better to certain class of drugs. It is useful to know which variants are prevalent in our population. Availability: Items available for loan: UVAS Library [Call number: 2939-T] (1).

61. Analysis Of Tp53 Gene Isolated From Oral Cancer Patients

by Amir Saleem (2013-VA-897) | Dr. Muhammad Wasim | Prof. Dr. Tahir Yaqub | Prof. Dr. Habib-ur-Rehman.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Cancer is a term used for disease in which abnormal cells divide without control and are able to invade other tissues. Cancer cells can spread to other parts of the body through blood and lymph system. TP53 is one of the most important tumor suppressor genes, mutated in more than 50% of human malignancies. It controls DNA repair, cell cycle and apoptosis and therefore plays an essential role in keeping genetic constancy. TP53 gene is present on the short arm of chromosome number 17. In human it extends 19,200 bp in 11 exons. Various parameters are used in the present study was aimed to investigate coding regions of TP53 gene for analyzing the mutations involved in oral cancer. Human OSCC samples (15) and normal tissue samples (15) were collected from Fatima Memorial Hospital Lahore. Samples were collected in tubes by oncologist containing ethanol and then brought to Molecular Biology and Biotechnology Lab of Institute of Biochemistry and Biotechnology, UVAS Lahore and frozen at -20°C before DNA extraction. Samples have been processed for research purpose. DNA was extracted from tissue by using Trizol Method and quantity was checked by nanodrop spectrophotometer. Two Primer sets were designed to amplify protein coding region of TP53 gene. After amplification through PCR, DNA Sequencing was done. Data interpretation was done by using several softwares like BLAST alignment tool, Bioedit, Clustal W2. In this study it was tried to find out the mutations in TP53 but no any kind of mutations were identified. Because I conducted my research only on 15 samples. So in future to use this gene as a potentional biomarker we can increase our number of samples. 46 The need for today is to develop valid biomarkers, which can be incorporated in ongoing in vivo and in vitro clinical mechanistic and improve the diagnosis and prognosis of this dreadful disease. In the conclusion, we must say that further study, with much larger sample number, is required to revise the effects of this polymorphism and accurately identifying the associated factors. There is a need to explore the other gene mutations causing oral cancer in local population of Punjab and Pakistan that will ultimately help to develop genetic counseling strategies, gene therapies and prenatal diagnostic procedures for the population of Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2937-T] (1).

62. Polymorphism Analysis Of Bovine Tumor Necrosis Factor Alpha Gene And Its Association With Mastitis In Nili Ravi Buffaloes

by Samia Tanveer (2011-VA-362) | Dr. Sehrish Firyal | Dr. Muhammad Wasim | Dr. Muhammad Nawaz.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Various number of factors cause hindrance in the milk production potential of buffalos. Mastitis is the costly and most prevalent disease causing production losses of dairy herds in Pakistan and elsewhere in the world. Susceptibility and resistance to mastitis is complex trait influenced by genetic variation of animals. Among these immunity gene variations, the polymorphism in tumor necrosis factor alpha gene (TNF-α) play important role in immune response to virus. Polymorphism in TNF-α gene is associated with mastitis susceptibility and resistance. It would be a potential candidate gene for imparting resistance mastitis in dairy buffalos. Blood sample were taken from the 20 Nili Ravi buffalos having clinical and subclinical mastitis. Extraction of DNA was done from frozen blood after thawing, using organic extraction method & also kit method followed by DNA quantification (i.e. gel electrophoresis and nanodrop). Total 5 primers were designed using Primer3 bioinformatics tool. All these primers were optimized using different protocols and a set recipe was obtained for each primer. The amplification of DNA samples was done one by one using all these five primers on optimized protocol. The amplicons obtained were subjected to agarose gel electrophoresis to check whether we have the required product or not using 100 kb ladder and then amplicones were send for the sequencing. Summary 110 The sequencing analysis of resulted amplicon sequence was done using Bioinformatics software Finch TV. Total of 6 mutations were found while 5 were same in all the samples whereas 6th mutation was found only in clinical samples. It is valuable in accomplishing genetic progress for resistance and to improve the immune response. This study will paved the way for animal breeder for selection of Nili Ravi mastitis resistant buffalos for breeding. TNF-α gene polymorphism based marker is now available for screening of resistant bulls as well. Availability: Items available for loan: UVAS Library [Call number: 2936-T] (1).

63. Genetic Identification And Molecular Classification Of Birds Commonly Found In Lahore Through Dna Barcoding

by Agha Wasif Ali Khan(2014-VA-01) | Dr.Ali Raza Awan | Dr.Muhammad Wasim.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2017Dissertation note: T Availability: Items available for loan: UVAS Library [Call number: 2938-T] (1).

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