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Home > Analysis of Cytochrome Oxidase-I Gene in Tilapia Fish Species Tilapia Zillii

Analysis of Cytochrome Oxidase-I Gene in Tilapia Fish Species Tilapia Zillii

Thesis Info

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Author

Ufaq Pervaiz

Institute

Virtual University of Pakistan

Institute Type

Public

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Software Engineering

Language

English

Link

http://vspace.vu.edu.pk/detail.aspx?id=305

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676721018868

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DNA barcoding is a technique concerned with the classification of things that is done with the help of small gene or DNA sequence with a known location on a chromosome that can be used to identify individuals or species in organisms mitochondrial DNA (mt DNA) which is helpful in recognition of meticulous species. It uses sequence variety in a 658-base pair fragment near the 5? end of the mitochondrial cytochrome c oxidase subunit I (COI) gene as a means for species recognition. DNA barcoding is a more defined and steady method in comparison with the classification based on the form and structure of the organism. It is equally useful in any stage of life cycle of fishes. The present research project was designed to recognize tilapia fish species (Tilapia zillii) of Pakistan genetically. Blood samples (n=30) were collected from Tilapia fish and genomic DNA was extracted and confirmed by 1% agarose gel. A short segment of COI gene (680bp) was amplified. PCR products were sequenced and analyzed by bioinformatics tools. Number of the haplotypes was 7. The haplotype diversity was Hd: 0.584 while nucleotide diversity was Pi: 0.00244. The mean intraspecific K2P genetic distance was 0.019. The estimated transition/transversion bias R was 1.40 that showed that this species possess very low genetic diversity. COI may supply a landmark for the classification of associated species at molecular level. As tilapia is extensively used for food and many other purposes so use of DNA barcoding technique is very helpful in discriminating it from other correlated fish species. It will also reduce the chance of mislabeling of tilapia fish species during its trading internationally as well as the species assessment at national level.
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مولانا عبدالمجید حریری

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Ethical and Moral Issues in Undergraduate Medical Education: An Exploratory Study

Background: Ethical and moral values form the basis of the environment in any academic institution. Objectives: This study aimed to explore the ethical and moral issues faced by medical students during their undergraduate medical education in Pakistan. Methods: This qualitative exploratory study was conducted at ABWA Medical College, Faisalabad. It included thirteen semi-structured, one-to-one interviews of volunteering undergraduate medical students of Punjab, Pakistan, from September 2021 to February 2022. The interview invitation was sent electronically to social media groups. Member checking of transcribed interviews was done, and data were analyzed thematically using Atlas-ti software by two independent researchers. Results: Thematic data analysis led to the formation of 28 codes, 13 subthemes, and 3 main themes. Based on the social constructive approach, three main themes were categorized into ethical issues in (1) academic, (2) social, and (3) clinical contexts. Conclusions: The results showed that undergraduate medical students face a myriad of ethical and moral disruptions during their undergraduate study period at medical college. Much awareness about identifying and dealing with such issues is recommended to bring more quality to the program.

Molecular Studies on Plant Based Cyclotides for Protein-Protein Interaction

Bioactive agents like secondary metabolites and peptides are gaining much interest while addressing the issues of agricultural and health threats including pests and pathogens and in drug development. Among the plant bioactive peptides, cyclotides being disulphide rich, stable, resistant and having ability to graft epitopes on it or allow sequence variations in its different loops that are very important regarding biological application and research interests for drug development and delivery. The present study was therefore focused on evaluating the bioactive potential of cyclotide bearing indigenous plants including Viola odorata, Viola tricolor, Viola hybrid, Petunia, Clitoria ternatea, pansy F1, Panicum vigatum, Panicum laxam, Panicum maximum and Hamelia patens. The extracts were prepared in phosphate buffer saline (PBS) and protein extraction buffer (PEB). Ptunia and MCOTI-I both separately showed highest DPPH activity (antioxidant) due toas the activity was reduced significantly after treating with Proteinase K. Hamelia possessed highest reducing power, hemolytic, thromobolytic and antimicrobial activites. Mutagenic responses of the (Ames test) medicinal plants were not significant. DNA demage protection assays were also done on all protein extracts including MCOTI-I. Cyclotide genes were also isolated from the selected plant’s DNA using specific primers. It was found that the chimeric arrangement of cliotide gene (from Clitoria plant) as most attractive with conserved sequences in cyclotide and Albumin-1 domains and little intronic variations. Moreover, for cyclotide-protein interaction studies, Texas Red-DBCO amine dye was synthesized to label MCOTI-I-AziF with p-azidophenylalanine Uaa in origami DE3. Optimizations were first done by using the MCOTI-I and its label form MCOTI-I-Lys-TxRd. The click reaction to bind TxRd-DBCO with MCOTI-I-AziF was finally done in buffered guianidinum HCl at a molar ratio of 1:100 and analysed through LC/MS and MCOTI-I-AziF-TxRd-DBCO was confirmed by mass spectrometry. For binding assay FRET analysis was done with Trypsin-EGFP, saturation was achieved with KD value 29.7 ± 1.08 nM. Optimization of same amount of MCOTI-I inteins (using westernblot) was taken in 3-4 h. The IPTG induction avoided the overnight incubations and unwanted backgrounds. The MCOTI-I a cyclotide was expressed for the first time in yeast cells. Current research opens new understandings towards the bioactive xii potential of cyclotide bearing plants with/without peptide content, genetics of cyclotides genes from indegenous plants, optimizations of expression and in vitro labeling studies of MCOTI-I for optical studies regarding drug development and targeting.