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Molecular Studies on Plant Based Cyclotides for Protein-Protein Interaction

Thesis Info

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Author

Mushtaq, Zahid

Program

PhD

Institute

University of Agriculture

City

Faisalabad

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016

Thesis Completion Status

Completed

Subject

Biochemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9619/1/Zahid_mushtaq_Molecular_Biology_2016_UOF_2016_29.11.2016.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726716199

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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.
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توفیق فاروقی

فاروقی، توفیق
بھائی توفیق فاروقی بھی اﷲ کو پیارے ہوگئے
موت توسب ہی کوآتی ہے اورآنی ہے ہی،جوچیز پیدا ہوئی ہے اسے فنا بھی ہونا ہے۔ موت بھی آنی ہے۔لیکن کچھ حضرات کی موت سے ایسا غم وصدمہ اور دکھ ہوتاہے کہ غم بھلانے سے بھی نہیں بھول پاتاہے۔
برہان میں وفیات کا صفحہ لکھاجاچکا تھا، شمیم عثمانی اور حضرت مولانا وحیدالزماں کیرانوی کے انتقال کی تعزیتی تحریر سپرد قلم کی جاچکی تھی کہ ابھی ابھی خبر ملی ہے کہ ’’خاتون مشرق ‘‘ اور ’’گلابی کرن‘‘ کے مدیر جناب توفیق فاروقی مورخہ ۲۴؍اپریل ۱۹۹۵ء بروز پیر کو عارضہ قلب میں مبتلا ہوکر انتقال فرماگئے ۔انا ﷲ وانا الیہ راجعون ۔
مرحوم توفیق فاروقی مجاہد آزادی مولانا عبداﷲ فاروقیؒ کے فرزند اکبر تھے، بڑے ہی ملنسار خلیق و متواضع اور یوپی ودہلی کی تہذیب ووضعداری کے نقیب و علمبردار تھے۔ انہوں نے ’’خاتون مشرق‘‘ اور ’’گلابی کرن‘‘ جیسے رسالوں کے ذریعے مشرقی خصوصاً مسلمان خاتون کو نیک وباعمل اورعفت وعصمت کاپیکر بنانے کا بڑا ہی اچھا اورقابل ستائش کام کیا۔
مفکر ملّت حضرت مفتی عتیق الرحمان عثمانیؒ سے مرحوم توفیق فاروقی اوران کے والد مرحوم مولانا عبداﷲ فاروقی نے ہمیشہ رہنمائی حاصل کی۔احقر عمیدالرحمن عثمانی سے بڑی محبت وعقیدت رکھتے تھے کہتے تھے کہ بھئی تم تو مجاہد ہوکہ اپنے ابّاجان اور ملّت کے رہبر مفتی صاحب کی یادگار ادارہ ندوۃ المصنفین اوررسالہ برہان کوقائم رکھاہوا ہے۔اﷲ تمہیں سلامت رکھے تمہارا دم بڑا غنیمت ہے۔
ذاتی معاملات میں مفتی صاحب کی وفات کے بعد بڑی مدد کی حمایت کی، بہرحال مرحوم توفیق فاروقی بڑے زندہ دل انسان تھے انہوں نے ادب و صحافت کی خدمت کی، ملک وملّت کی خدمت کی۔مسلمان عورت کواسلامی خاتون بنانے کے کاموں کوبڑی اچھی طرح انجام دیا۔اﷲ تعالیٰ غریق رحمت فرمائے اٰمین۔
ادارہ ندوۃ المصنفین اوررسالہ برہان کی طرف سے احقر عمید الرحمن...

الحوكمة المؤسسية بمدارس دولة جنوب أفريقيا وإمكانية الإفادة منها بسلطنة عُمان

هدفت الدراسة الحالية إلى التعرف على الحوكمة المؤسسية بمدارس دولة جنوب أفريقيا وإمكانية الإفادة منها بسلطنة عُمان، واتبعت الدراسة المنهج الوصفي، كما استخدمت نظرية تحليل المضمون في تحليل الوثائق في جمع البيانات والمعلومات. وتوصلت الدراسة إلى مجموعة من النتائج أهمها: حاجة المدارس في سلطنة عُمان إلى هيئة مُحددة تتولى مسؤولية الحوكمة المؤسسية في المدارس، وتضم هذه الهيئة ممثلين عن كافة المُشاركين والمستفيدين من العملية التعليمية من إدارة مدرسية ومعلمين وأخصائيين وطلبة وأولياء أمور وأعضاء مجتمع محلي، ويتم تدريب هذه الهيئة على القيام بواجبات ومسؤوليات وأدوار مُحددة، وقيام هذه الهيئة بتشكيل مجموعة من اللجان المتنوعة لدعم عملها

Comparative Performance of the Students of Higher Secondary Schools and College at Intermediate Level in Punjab

The main purpose of the study was to evaluate the overall performance of the students of higher secondary schools and colleges at intermediate level in Punjab. Higher secondary school level is the intermediate stage, where students have to carry out their future planning. It is necessary to improve the academic performance of students at higher secondary level of education, particularly in the field of Mathematics because the mathematics curriculum identifies the different attributes of learning which is to be assessed through the performance of students. Therefore the study was conducted to evaluate the performance of the mathematics students of HSSs and colleges at intermediate level. The major objectives of the study were to analyze overall performance of the students at intermediate level in higher secondary schools and colleges in a comparative perspective; to investigate factors affecting teaching-learning environment faced by the students and teachers in Higher Secondary Schools and Colleges of Punjab; to evaluate performance of male and female students of Mathematics in Higher Secondary Schools and Colleges and to explore performance of rural and urban area students of Mathematics. The population of the study comprised 946 principals, 1336 maths teachers and 73455 maths students of higher secondary schools and colleges of Punjab province. Province of Punjab is densely populated and consists of 37 districts. These districts are administratively divided into nine divisions. Each division has a BISE (Board of Intermediate and Secondary Education). Keeping in view the short span of time and resources constraints, the study was delimited to two Boards, i-e Rawalpindi and Sargodha. For evaluation of the performance of the students in comparative perspective, the research was also delimited the results of the students of the Mathematics part-1 at intermediate level in higher secondary schools and colleges. The researcher used three questionnaires (First for the principals, second for the mathematics teachers and third for the students of mathematics part-2) as research instruments. The researcher personally visited the selected institutions and the questionnaires were administered to 440 mathematics students of part-2 at intermediate level, 60 mathematics teachers and 48 principals of Government higher secondary schools and Government colleges. The respondents were requested to give their responses to each item on three-point Likert Rating Scale. The researcher also visited BISE Rawalpindi and BISE Sargodha for obtaining HSSC-I Annual Examination 2012 results of the students included in the sample. The data were analyzed by using SPSS version-21 and statistical techniques (ttest, chi-square-test “Monto Carlo Simulation Method”). Themain conclusions of the study were: 1. Overall performance of the students of HSSs’ and colleges was indifferent. However, performance of higher secondary schools students of Rawalpindi Board was better than those of Sargodha Board. In colleges, the performance of the students of Sargodha Board was better than those of Rawalpindi Board. 2. The performance of students of Rural and Urban institutions was similar both higher secondary schools as well as for colleges. 3. The performance of boys and girls was significantly different. The performance of girls was better than performance of boys. 4. The math’s teachers were aware of the objectives of teaching Mathematics at higher secondary level who completed syllabus with in time. In both setups, majority of the teachers used demonstration, inductive and activity based methods. Mostly teachers of HSS were given individual attention and used appropriate teaching aids effectively to the students as compare to colleges. 5. Mostly teachers of HSS’s gave individual attention to students who used appropriate teaching aids effectively to the students as compared to colleges.