http://20.198.91.3:8080/jspui/handle/123456789/145 2026-03-09T04:06:58Z http://20.198.91.3:8080/jspui/handle/123456789/8580 Title: GM2 mediated YAP/TAZ regulation: elucidation of critical involvement of the master regulator LATS kinases Authors: Kar, Avinandan Abstract: Hippo signaling is an evolutionarily conserved regulator of tissue growth and cell fate. LATS1/2 is the upstream kinase of the Hippo signaling pathway which regulates phosphorylation and dephosphorylation of YAP/TAZ to modulate nuclear-cytoplasmic shuttling. To define the role of GM2 in regulation of Hippo pathway, here we aimed to generate LATS2 knockout cell line. CRISPR-Cas9 genome engineering technology has been applied to achieve this goal. Here, we have designed and constructed CRISPR-Cas9 plasmid against coding sequence of LATS2. The sequence verified CRISPR construct against human LATS2 were then transfected into HeLa cells to assess the functionality and to generate LATS2 knockout Hela cell Line. Further, LATS2 CRISPR construct transfected Hela cells were selected and clonal selection strategy was undertaken to select, expand and clonally purify LATS2 knockout Hela cells. Here, we have shown that CRISPR-Cas9 construct against LATS2 is functional and we were also able to generate LATS2 knockout Hela cell line. This LATS2 knockout cell will help us to understand the role GM2 in regulation of Hippo signaling in future studies. 2019-01-01T00:00:00Z http://20.198.91.3:8080/jspui/handle/123456789/8579 Title: Designing and construction of a mammalian expression cassette containing monomeric green fluorescence protein gene in pCDNA3.1 by molecular cloning method Authors: Ghosh, Kripanjali Abstract: Green fluorescence protein (GFP) is a small but a very stable protein which absorbs blue light and fluoresces green. It has a very negligible toxicity to the cell it expresses. Using these characteristics, protein tagging to the N or C terminus of GFP has revolutionized the research field. It has a wide range of application and various advantages in cell biological and proteomics study. Here in this dissertation we have successfully cloned the GFP gene within mammalian constitutive expression vector pcDNA3.1 which can be used to tag any protein of interest to the N or C terminus of GFP to visualize its localization and its interaction with other proteins through confocal microscope or any other fluorescence microscope. 2019-01-01T00:00:00Z http://20.198.91.3:8080/jspui/handle/123456789/8578 Title: Studies on purification of ΒC1 protein of geminivirus satellite and geminivirus pathogenesis Authors: Ghosh, Ananya Abstract: Begomoviruses constitute the largest genus of the family Geminiviridae which causes huge loss of the crops worldwide. Monopartite begomoviruses are usually associated with betasatellite which highly increases disease development process and therefore possess a great threat for the economically important crops. βC1, the only protein encoded by the betasatellite plays multitasking roles and aid helper virus to promote infection by increasing symptom severity, by suppressing the transcriptional and post transcriptional gene silencing etc. In this study PatnaβC1 protein encoded by Tomato leaf curl Patna betasatellite and its ATPase deficient mutant has been purified by affinity chromatography to examine the structural changes of the protein after the mutation. Furthermore to study the begomovirus mediated pathogenesis, Nicotiana benthamiana plants infected with Tomato leaf curl New Delhi virus DNA-A (ToLCNDV-A) and its mutants along with Radish leaf curl betasatellite (RaLCB). The plants infected with ToLCND-A along with RaLCB shows severe symptoms like veinal chlorosis, leaf curling, stunted growth etc. However plants infected with ToLCNDV-AC2 and ToLCNDV-AC4 along with RaLCB did not shows any symptoms. RaLCB specific PCR analysis did not support the result observed for ToLCNDV-AC4 – RaLCB inoculated plants, however the PCR analysis of ToLCNDV-AC2 – RaLCB inoculated plants suggests the influence of RaLCB for symptom development. 2019-01-01T00:00:00Z http://20.198.91.3:8080/jspui/handle/123456789/8577 Title: A study on the synthesis of Bi-metallic copper-silver nanoparticle & its anti-bacterial potency against multi-drug resistant diarrhea-causing E.coli ETEC in vitro and in vivo in mice Authors: Kundu, Subhajit Abstract: The development of easy methods of preparation metallic nanoparticles (NP) has been very much attracting to the field of material science, from their application potential in different areas, due to their unusual size-dependent optical and electronic properties. This study deals with the development of a simple robust method of synthesis of simple bimetallic copper-silver nanoparticles (Cu-AgNP) by successive reduction of Cu (No3)2 and AgNo3, using hydrazine hydride as the reducing agent and gelatine and poly-vinyl pyrrolidone (PVP) as the capping agents (stabilizer). The round shaped particles were of core-shell structure with a core of Cu0 atoms surrounded by a shell of Ag0 atoms. The size and Zeta potential of the NPs were (76.26) nm and (-15.5) mV respectively. The particles were crystalline in nature and 90% of the precursors Cu (No3)2and AgNo3 were converted to Cu-Ag NPs. The NPs were characterized by the techniques like dynamic light scattering (DLS), atomic force microscopy (AFM) etc. The NPs were toxic to multi-drug (cefixime, ceftriaxone, erythromycin, sulfamthoxazole, trimetthoprin, vancomycin, tetracycline, rifaximin etc.) resistant Enterotoxigenic Bacteria (ETEC) that killed most of the ETEC. Therefore, Cu-Ag NPs can be used as a potent antibacterial drug in future. The antibacterial activity of this Cu-Ag NPs on Gram negative ETEC was demonstrated by agar plating method. Determination of the minimum inhibitory concentration (5.483μg/ml), minimum bacterial concentration (6.27μg/ml), showed that our Cu-Ag NPs were highly effective at lower concentration than that of any antibiotic. 2019-01-01T00:00:00Z