1.
Role Of Maoa Polymorphism In Criminal Violence Among Convicted Offenders
by Shahpal Shujat (2010-VA-494) | Dr. Maryam Javed | Dr. Wasim Shehzad | Dr. Saadat Ali.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Criminal violence is the violent act of crime that can be of different category such as Homicidal, Sexual and Physical violence. Criminal violent behaviour is under the control of brain having serotonergic and dopaminergic system that is under the influence of genes .MAOA gene lies under the control of serotonergic system. There are 2 polymorphic forms of MAOA gene on the basis of its activity i-e MAOA-L (Low activity allele) and MAOA-H (High activity allele). MAOA-L in males but in females MAOA-H is linked with violent behaviour in combination with environmental factors.
Blood/Saliva /Buccal swabs samples (n = 20) were collected from District Jail Sheikhupura, Pakistan Organic/Inorganic method of DNA extraction was used. Primers for PCR amplification was designed using Primer3 software. PCR products were sequenced by using Sanger method from CAMB. Violent behaviour was determined by using Buss and Perry aggression Questionare Physical aggression scoring.
Sequencing results were analyzed using CHROMAS software. Sequence alignment tool like CLUSTAL was used for aligning multiple sequences of convicted samples and compared with control (DNA mixture amplification) and standard samples and detected for SNP. Then SNP were detected for the amino acid change by using ExPASy software.SNP statistical analysis was done by calculating POPGENE software and association analysis was performed by using ANOVA.
SNP in exon 8 at locus 43591035 of MAOA was identified that was T instead of G while in exon 13 two SNPs were identified at locus 43603089 and 43603112 .Both SNPs for exon 13 was heterozygous and changes T into A .The synymous SNPs were at locus 43591035 and
Summary
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43603089 . But the SNP at locus 43603112 was non-synonymous .The association study showed that there was no association between SNP and violence scores among convicted offenders. Availability: Items available for loan: UVAS Library [Call number: 2486-T] (1).
2.
Genetic Study Of Slc6a4 Gene In Convicted Offenders From Prisons Of Punjab, Pakistan Exhibiting Antisocial Personality Disorder Traits
by Asima Saman | Dr. Saadat Ali | Dr. M. Yasir Zahoor | Dr. M. Imran.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Antisocial Personality Disorder (ASPD) is characterized by incapacity of an individual to adapt themselves to social norms. Patients with ASPD typically have irritability problems and aggressive feelings toward other people. The serotonin transporter gene (5-HTT orSLC6A4) has been associated with regulation of serotonergic neurotransmission, mood and behaviour traits.Low levels of the neurotransmitter serotonin are believed to affect judgement, planning, and impulse control in ASPD sufferers. Genetic polymorphism in selected intronic region of serotonin transporter SLC6A4 gene was associated with antisocial personality disorder (ASPD) in convicted criminal of Punjab, Pakistan. We have selected two regions from SLC6A4 gene, which was intron 1 and exon 3.After extraction of DNA Polymerase Chain Reaction was used to amplify the extracted DNA of criminals (n=20) and control (n=10) for selected segments of intron 1 and exon 3 of SLC6A4gene. Sanger’s DNA sequencing method (di-deoxy chain termination method) was used to sequence the amplified fragments. Statistical and bioinformatics tools were used to analyse the data. Intron 1 has shown 5-HTTLPR polymorphism S allele (0.85 frequencies), LA allele (0.05 frequencies) and LG allele (0.1 frequencies) and exon 3 did not show polymorphism in criminal’s sample. The study highlights the role of SLC6A4 gene polymorphism in criminals of Punjab having antisocial personality disordertraits. Availability: Items available for loan: UVAS Library [Call number: 2576-T] (1).
3.
Sequence Analysis Of Violent Behavior Gene Among Criminals
by Jawairia Akram (2010-VA-492) | Dr. Asif Nadeem | Dr. Muhammad Imran | Dr. Saadat Ali.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Violence is defined as uncontrolled emotions problem and is a reason of violent behavior among criminals. Violence is mostly physical towards other people. MAOA and MAOB are isozymes of monoamine oxidase. MAOA is associated with aggression and violence in criminals as it affects brain structure and function which ultimately causes violence and aggression MAOA gene present on mitochondrial outer membrane encodes monoamine oxidase that degrade neurotransmitters like dopamine, serotonin, epinephrine and nor epinephrine. An SNP (MAOA-LPR) in long promoter region of MAOA alters transcriptional activity of monoamine oxidase A and have two allelic forms MAOA-L and MAOA-H. MAOA-L is low activity allele and MAOA-H is high activity allele. Different research study suggested that MAOA-L is strongly associated with criminal activity in males. Aim of the study was to analyze the sequence of extreme violent behavior gene (MAOA) among criminals. Samples (n= 20) were collected from convicted offenders. Control samples (n=20) were collected from UVAS students. Organic method of DNA extraction was used. BPAQ (Buss and perry aggression questionnaire) was also filled by all the subjects included in the study. Primers for PCR amplification were designed using Primer3 software. PCR products were sequenced bi-directionally on ABI 3130XL Genetic analyzer. Results of sequencing were analyzed using CHROMAS software. Sequence alignment tool like BLAST (Basic local alignment search tool) was used for SNPs identification. 3 intronic and 1 exonic SNPs were observed and confirmed by BLAST. Exonic SNP gave significant p values computed by Chi square calculator. However, intronic SNPs were not significant according to chi square test. SNPs identified were not found to be associated with self-reported aggression. SNP observed in exon 14 is reported to be involved in psychiatric and depressive disorders. Availability: Items available for loan: UVAS Library [Call number: 2566-T] (1).
4.
Assessment Ofgenetic Polymorphism In The Tph Gene As Susceptible Factor For Aggressive Behavior In Criminals From Prisonsof Punjab, Pakistan
by Zonash Riaz (2010-VA-479) | Dr. Saadat Ali | Dr. Muhammad Yasir Zahoor | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Aggression is perceived as hostile, injurious, or destructive behavior often caused by frustration, can be collective or individual. Genetic studies have associated several genes with aggression in humans. One of the candidate genes that turned out to be associated with aggression, anger, and impulsivity is the tryptophan hydroxylase (TPH) gene. We investigated the polymorphism in the TPH gene in the unrelated male individuals in the Punjab ethnic backgrounds who were administered the Punjabi translation of Buss and Perry aggression questionnaire. The questionnaire measured four aspects of aggression: physical aggression, verbal aggression, anger and hostility (Buss and Perry, 1992).Scores ± SD of 83.544± 26.63 was obtained for Buss and Perry aggression questionnaire. TPH is a rate-limiting biosynthetic enzyme in the serotonin pathway and regulates levels of 5-hydroxytryptophan (5-HT) by converting tryptophan into 5-hydroxytryptophan, which is the direct precursor of 5-HT.
It is conceivable that variations in the TPH gene could contribute to low activity of the 5-HT system. Single nucleotide polymorphisms (SNPs) that show associations to aggression and anger-related traits have been detected in intron 7 of TPH gene.DNA of individuals categorized into controls and criminal groups was extracted by organic method of DNA extraction.The targeted region of the TPH gene was amplified by the primers designed against intron seven. The amplified Pcr product was precipitated and it was sent for sequencing. The resultant sequenced data was then compared on the basis of Buss and Perry aggression scores. All unrelated male individuals from the Punjab ethnic groups were assessed on the scales showing scores for physical aggression, verbal aggression, anger and hostility. The minimum score for the respondents were 65 and highest score for the respondents were 135 among the criminal group while control have minimum scores of 50 and maximum scores of 113.Mean scores and standard deviations were calculated for criminals and control groups. Control group havephysical aggressionmean scores ± SD19.318 ± 6.21, verbal aggressionmean scores ± SD17.590± 4.41,angermean scores ± SD23 ± 6.868and hostility mean scores ± SD23.636± 9.12and total mean scores ± SD83.544± 26.63while criminals have physical aggressionmean scores ± SD28.2±8.134, verbal aggressionmean scores ± SD20.4±4.427, anger mean scores ± SD27.3±6.97and hostilitymean scores ± SD28.1±7.72and totalmean scores ± SD04.2±20.47.Mean aggression values for the criminals was 104 and for controls was 83, higher in criminals as expected. Criminals groups exhibited greater level of aggression as compared to that of control groups on the basis of four scales of aggression i.e. physical aggression, verbal aggression, anger and hostility.
Observed genotypic frequencies among the control groups were 0.7 for CC, 0.3 for the AC and 0 for AA whereas genotypic frequencies amongst criminal group were 0.3 for CC, 0.6 for AC and 0.1 for AA. Controls carried higher genotypic frequencies for normal CC genotype than criminals whereas the genotypic frequencies for AA and AC genotypes were higher in Criminal group.Observed allelic frequencies amongst the control group was 0.8 for C and 0.15 for A whereas observed allelic frequencies amongst the criminal group was 0.4 for A and 0.6 for C. Controls carried higher allelic frequencies for the normal C allele while criminals carried higher allelic frequencies for A allele.In our study proportion of the less common (A or U) alleles was 40%, and the proportion of the more common (C or L) alleles was 60% in criminal group as compared to 15% of A allele and 85% of C allele in the control group. Statistical analysis has associated significantly Criminals and controls group at P value less than 0.05.
Advances in the understanding of the genes modulating aggression can contribute meaningfully to a rational assessment and treatment of individuals with pathological aggression and a predisposition to violence. Results can be utilized for the screening of Aggression in the individuals for forensic applications. In future studies, other polymorphism in TPH and other aggression related genes may also be analysed in Pakistani population.
Availability: Items available for loan: UVAS Library [Call number: 2595-T] (1).
5.
Genetic Analysis Of Slc24a5 Polymorphism In Pakistani Population, In Association With Human Skin Pigmentation As An Externally Visible Characteristic Parameter
by Asma Hameed (2008-VA-332) | Dr. Saadat Ali | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Imran.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Human skin pigmentation is a phenotypic trait that varies within a population or among different populations. In addition to the genetic factors, some of the diseases (may be genetic or epigenetic), exposure to UV or usage of cosmetics may also be involved in the pigmentation outlook. It is possible to predict human identity on the basis of DNA polymorphisms in the genes coding human phenotypic characteristics.
In case of human skin pigmentation various genes are responsible to code variability among which SLC24A5 is an important contributor. This area of research is important in the field of forensic science in cases where reference samples are not available for comparison with the DNA profiles obtained from the crime scene evidence. SNPs in the coding region of exon3 (84bp) of SLC24A5 related to skin pigmentation (as reported in literature) are associated to a predictable variation in skin color in Pakistani Population.
Blood samples (62) were collected from the participants having three types of skin coloration fair= 20, medium=22 and dark=20 from general population belonging to Punjab. Organic method (Phenol chloroform extraction method) of DNA extraction was used. After extraction DNA was quantified on nanodrop spectrophotometer. Primers for the exonic region 3 of SLC24A5 gene were designed using primer 3 software. PCR amplification of the selected region was done through touch down PCR. DNA after obtaining PCR products was purified and the samples were sequenced bi directionally on ABI 3130XL Genetic analyzer.
The results of sequencing were analyzed using CHROMAS Lite 2.1 software. Sequence was converted into Fasta Format required for alignment study. Alignment tools like Blast were required for SNPs identification and comparison of all the sample sequences with the reference sequence. Mean color scores and mean ages of all the skin color groups were calculated separately in both male and female participants. Two types of genotypes were observed i.e, AA and AG. 24 out of the total sample size showed heterozygous peaks and confirmed the polymorphism also in Pakistani population at position 299 of the sequence. Difference between allelic and genotype frequency of studied gene were evaluated and by t test and association analysis to check out the significance of the studied data with the skin coloration was done and it was concluded that AA genotype is significantly associated with fair skin color in male and female population. Furthermore, AG genotype was significantly associated with dark skin coloration in female population. This type of study reveals that after the genetic analysis of the DNA obtained from the crime scene, prediction of skin color/hue of crime related individuals of fair skin color as well as dark skin color belonging to Pakistani Population can be made in those cases where reference samples are not available. So this can be used as a genotypic marker for screening out and forensic identification of individuals in various crime cases where reference samples are not available for comparison purposes and matching suspects. Availability: Items available for loan: UVAS Library [Call number: 2608-T] (1).
6.
Molecular Characterization Of Oca2 Gene In Correlation With Eye Color For Forensic Application
by Anam Noor (2014-VA-942) | Dr. Muhammad Yasir Zahoor | Dr. Allah Rakha | Dr. Saadat Ali | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: DNA phenotyping is the use of genetic information such as DNA to determine a phenotype. It helps forensic investigator to predict the physical appearance of an individual to find unknown perpetrators or to identify missing persons using molecular analyses from biological samples in cases where all other means of inquiry, including conventional DNA profiling are non-informative. In a non-forensic setting, it permits the prediction of the physical appearance of our ancestors, historical persons or any other deceased individual for whom the identification of appearance traits may be interesting, and it sheds light on human evolution. Based on current research there are only a few traits for which it is possible to make an accurate description based on underlying genetic variation.
Eye color is a complex polygenic trait and is under the control of many genes. There are infinite number of eye colors with a multitude of patterns and mixtures. Almost 74% human eye color is under the control of OCA2 gene on chromosome 15. This gene correlates with the physical appearance of eye color as EVCs (externally visible characteristics) therefore it can be used as a parameter in forensic application.
Samples collected from local areas of Pakistan is divided into two groups brown that include samples from 17 individuals and other than brown including 15 individuals. DNA of 32 samples was extracted and samples were amplified against a selected sequence of OCA2 gene containing SNP rs1800407, which was previously reported to be associated with eye color in European populations. These amplicons were sequenced using Sanger sequencing and chromatograms obtained were analyzed by pairwise and multiple alignment tools. The results show the presence of5 polymorphic sitesin various samples including SNPsrs1800407 and rs1900758. These polymorphic sites were further analyzed by applying t-test which shows no significant association between retrieved polymorphic sites and eye color.The results show no significantly associated marker with eye color to be present within the selected sequence so we need to analyze other markers or SNPs which could be found to be associated with eye color that would be very useful in forensics application.
Availability: Items available for loan: UVAS Library [Call number: 2623-T] (1).
7.
Development Of Novel mtDNA Metabarcodes For Species Differentiation Of Class Mammalia
by Rabia Latif (2014-VA-952) | Dr. Muhammad Imran | Dr. Saadat Ali | Dr. Akhtar Ali.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation 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 Vrtebrata such as Class Mammalia. 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 Mammals for different forensic and molecular biodiversity analyses.
Mitochondrion, the energy coins for the cell, performs the function of the oxidative phosphorylation and the formation of ATP also called energy coins for the cell. Mammalian mitochondrial genome (mtDNA) is a double stranded, circular, covalently closed molecule of approximately size of 16.4 kb. The mtDNA is inherited from the mother as a haploid and heteroplasmy has been found hardly.This fact makes it potentially relevant in the identification of maternal relationships, absence of recombination and the fast rate of evolution Blood/tissue samples were collected from Class Mammals (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 mammalian 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 Analyzed following Sanger’s dideoxy method of
Summary
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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 the origin of unknown mtDNA sequences. Both sequencing experiments and phylogenetic studies confirmed the specificity of the universal primer set developed and present a novel metabarcode found in this region of genome (16SrRNA) for species level identification of large number of mammalian species. 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: 2618-T] (1).
8.
Microbiome Analysis Of Human Normal Specific Flora From Skin Of Laborers And Academic Professionals Of Lahore For Forensic Application
by Talha Umair (2014-VA-941) | Dr. Wasim Shehzad | Dr. Saadat Ali | Dr. Muhammad Yasir Zahoor.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Human microbiota or normal flora is the aggregate of microorganisms that resides on the
surface of skin, oral mucosa, conjunctiva and GIT. Human skin has a complex variety of
microbial system and varieties of microbes mean that they are potential source of forensic
identification because human microbiome varies individual to individual due to differences
in hygiene, professions and region to region because of some environmental factors and
microbial flora can shed more frequently upon touching any kind of surfaces and microbes
are left for long time at any surface so can be identified easily.
Human microbiota varies individual to individual so it may become potential source for
forensic identification of individuals through specific microbiome analysis.
Fourty Samples were obtained by swabbing from the palm surfaces of hands and soles of feet
of individuals of different professional groups in order to recover bacterial communities.
Bacterial culturing and Bacterial DNA extraction followed by the implementation of 16S
rRNA amplification by polymerase chain reaction (PCR) and sequencing of the PCR
product, allowed an even more comprehensive broad range investigation of bacterial
communities. Bioinformatics analysis was done to compare microbial communities.
This research elaborated the significance of skin microbial communities in identifying
individuals and can be a major contribution in forensic science to find and identify
individuals when there is less major evidence, i.e. human DNA and body fluids. Availability: Items available for loan: UVAS Library [Call number: 2639-T] (1).
9.
Development Of Novel MtDNA Metabarcodes For Species Differentiation Of Class Reptilia
by Imran Tariq (2014-VA-505) | Dr. Muhammad Imran | Dr. Wasim Shehzad | Dr. Saadat Ali.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The FoImer COI: mtDNA. universal primers that are considered standard for DNA barcoding of life contain so many =matches against the target sequences of vertebrate men tat they often end in failure to amplify many of vertebrate DNA eurections. This disaepancy fawn foe the seaman and designing of new metabarcode panes that can be used m ideally all inditdrals of vertebrees or at least all individuals represented in a class of Vertebrate such as Cass Reprilia. The current study was embadang on such an endeavor
The proposed study was develop new m5DNA membarc ode that may be used as universal Kilns; to amplify almost all species of Class Repalia for different formic and molectdr biodivesity analyses.
Blood and tissue samples were collected from Class Repdha (at :east 24 species from every ceder reported to be present in Pakistan) DNA was extracted from the collected specimen through stacdasd organic method. qualified and =meted and then PCR-amplified using novel universal primers selected from aligned =DNA sequences origtadng from all repdlian mitochondria DNA pnomes submitted to diens online sequence databases such as NCB: micleotide database. Tne sensitiviry. of PCR was assessed using a range of DNA come:madam. The amplified products were sequenced on A131 Genetic Analyzed following Sarge's dideacy method of sequencing.
The correctness of obtained croDNA sequences were examined visually in Chromes Lite 2.1 software and then alipmmt of these sequences were per: waxed agitinc highly similar DNA sequences in NCBI nu6eonde databases using BLAST in order to identify the coigin of la-noun =DNA sequences
sequencing everimeas and phyla...net< studies was confirm the specificity of the universal primer set developed and present a novel metabarcode for species level identification of large number of reptelian species. So, In future this barcode can be used for species identification in various fields of study such as illegal trade and molecular estimation of boidiversity.
Availability: Items available for loan: UVAS Library [Call number: 2628-T] (1).
10.
Homology & Polymorphism Analysis Of Cc2d1a Gene In Human And Canine For Cognitive Function
by Hafiz Qamar Abbas (2014-VA-214) | Dr. Muhammad Yasir Zahoor | Dr. Wasim Shehzad | Dr. Saadat Ali.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Cognitive disability is a group of genetically heterogeneous abnormality that leads to variable degrees of cognition deficits. It has been shown that inherited disorders can be caused by mutations in large number of different genes and there is evidence for the presence of as yet unknown genes in a significant proportion of patients. This disease can affect 1-3% of overall population and higher in consanguineous families. We aimed to identifying the homology and polymorphism of the gene CC2D1A between human and canines. The present research work was carried out in four phases. The first phase was including enrolment of 10 affected non relevant families with disease history and consent was taken on consent forms as approved by IRB, UVAS. Secondly DNA extraction was done by using standard lab protocols. Thirdly amplification of the selected domains of selected gene (CC2D1A) was done through PCR amplification after designing primers of the selected domains. Sequencing of the amplified products has to be done through Sanger method and mutation analysis was conducted for variants We found two new asynonymous mutation one is deletion of c. 1664_1664delA which lead to the change in the normal function of protein (88%) and other is heterozygous mutation c.1921A/T that result in amino acid change from R to W (12%). Whereas homology analysis shows that deletion region is partially conserved as it code different amino acid but some key domains are conserved. This homology shows that deletion in this region can change the protein expression which can relate to unconscious condition like behavioral or mental retardation. This will be helpful in providing genetic counseling services to indigenous population for intellectual disability cases. Availability: Items available for loan: UVAS Library [Call number: 2627-T] (1).
11.
Eye Color Prediction Using Snps Of Oca2 And Herc2 Genes In Different Eye Color Groups From Pakistani Population
by Iqra Baig (2015-VA-813) | Dr. Muhammad Yasir Zahoor | Dr. Saadat Ali | Dr. Amjad Riaz.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2017Dissertation note: The outward appearance of living organism that can be visualized is known as External visible characteristic (EVCs). These are related to the interaction of different genes among themselves and with their environment. Through this technique police investigators or other forensic investigators determine perpetrators which are completely unknown to investigating authourities or to pinpoint missing persons utilizing biological samples in those situations where all other evidences of query, along with conventional DNA profiling give non-uniformities. Eye color is a multiplex trait controlled by many genes but the two major genes which play crucial role to determine eye color are OCA2 and HERC2 genes. 74% eye color of human is under the control of OCA2 gene and its function is influenced by HERC2 gene. These are present on chromosome 15. Eye color trait has miscellaneous inheritance pattern which does not obey simple pattern of Mendelian inheritance.
Blood samples of 40 volunteers along with photographs of iris collected from local population of Pakistan by categorizing different eye colors. DNA was extracted using organic extraction method. Then amplified using PCR with primers of SNP rs1800401 of OCA2 gene and SNP rs12913832 of HERC2 gene. Primers were designed through primer 3 software. Amplicons were analyzed by gel electrophoresis. Samples were sequenced by Sanger sequencing and chromatograms were analyzed by pairwise and multiple alignment tools.
We mapped five polymorphic sites in OCA2 gene including SNPs rs1800401 and rs10300271. Polymorphic sites of OCA2 are 89406 C>T, 89401 G>T, 894019 A>T, 89422 A>C and 89435 C>A. Six polymorphic sites of HERC2 also analyzed including SNP rs12913832 at 206678 T>C and other polymorphic sites are 206617delA, 206631delA, 206683 T>A, 206688delA and 206713delA. These polymorphic sites were further analyzed by applying t-test which shows no significant association between retrieved polymorphic sites and eye color except polymorphic site 89422 with genotype A>C in OCA2 gene (novel) and polymorphic site 206678 with genotype T>C in HERC2 gene (already reported) both are associated with non-brown eye color variation in our study. In conclusion, more research to DNA based human appearance prediction is recommended using large sample size as there are more SNPs also involve that would be very useful for identification or investigative leads in forensic future aspects.
Availability: Items available for loan: UVAS Library [Call number: 2902-T] (1).
12.
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: ; 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).
13.
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: ; 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).
